65 results on '"SULT2B1"'
Search Results
2. Knockdown of sulfotransferase 2B1 suppresses cell migration, invasion and promotes apoptosis in ovarian carcinoma cells via targeting annexin A9.
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Gao, Haocheng, Xia, Mengjuan, and Ruan, Heqiu
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CELL migration , *WOUND healing , *FLOW cytometry , *CANCER invasiveness , *APOPTOSIS , *OVARIAN tumors , *CELL proliferation , *CELL motility , *REVERSE transcriptase polymerase chain reaction , *CELL lines , *GENE expression , *BIOINFORMATICS , *METASTASIS , *MICROBIOLOGICAL assay , *TRANSFERASES - Abstract
Background: Sulfotransferase family 2B member 1 (SULT2B1) has been reported to play oncogenic role in many types of cancers. Nevertheless, the role that SULT2B1 played in ovarian cancer (OC) and the hidden molecular mechanism is obscure. Methods: Expression of SULT2B1 in OC was analyzed by GEPIA database. qRT‐PCR and western blot (WB) was applied for the appraisement of SULT2B1 and Annexin A9 (ANXA9) in OC cell lines. The capabilities of cells to proliferate, migrate and invade were assessed with CCK‐8 assay, wound healing assay, along with transwell assay. Cell apoptotic level was estimated utilizing flow cytometry. WB was employed for the evaluation of migration‐ and apoptosis‐related proteins. Bioinformatic analysis and co‐immunoprecipitation were used to predict and verify the combination of SULT2B1 and ANXA9. Results: The data showed that SULT2B1 and ANXA9 were upregulated in OC cells. SULT2B1 depletion suppressed the proliferative, migrative, and invasive capabilities of SKOV3 cells but facilitated the cell apoptosis. SULT2B1‐regulated ANXA9 expression and were proved to bind to ANXA9. Additionally, ANXA9 deficiency exhibited the same impacts on cell migrative, invasive capability and apoptotic level as SULT2B1 silencing. Moreover, ANXA9 overexpression reversed the inhibitory impacts of SULT2B1 silencing on the proliferative, migrative, invasive, and apoptotic capabilities of SKOV3 cells. Conclusion: In summary, SULT2B1 silencing repressed OC progression by targeting ANXA9. [ABSTRACT FROM AUTHOR]
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- 2024
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3. Human sulfotransferase SULT2B1 physiological role and the impact of genetic polymorphism on enzyme activity and pathological conditions
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Fatemah A. Alherz
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cholesterol sulfotransferase ,SULT2B1 ,autosomal recessive ichthyosis ,polymorphism ,pregnenolone sulfotransferase ,colon cancer ,Genetics ,QH426-470 - Abstract
Human SULT2B1gene is responsible for expressing SULT2B1a and SULT2B1b enzymes, which are phase II metabolizing enzymes known as pregnenolone and cholesterol sulfotransferase (SULT), respectively. They are expressed in several tissues and contribute to steroids and hydroxysteroids homeostasis. Genetic variation of the SULT2B1 is reported to be associated with various pathological conditions, including autosomal recessive ichthyosis, cardiovascular disease, and different types of cancers. Understanding the pathological impact of SULT2B1 genetic polymorphisms in the human body is crucial to incorporating these findings in evaluating clinical conditions or improving therapeutic efficacy. Therefore, this paper summarized the most relevant reported studies concerning SULT2B1 expression, tissue distribution, substrates, and reported genetic polymorphisms and their mechanisms in enzyme activity and pathological conditions.
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- 2024
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4. Sulfotransferase SULT2B1 facilitates colon cancer metastasis by promoting SCD1‐mediated lipid metabolism.
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Che, Gang, Wang, Wankun, Wang, Jiawei, He, Cheng, Yin, Jie, Chen, Zhendong, He, Chao, Wang, Xujing, Yang, Yan, and Liu, Jian
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LIPID metabolism , *COLON cancer , *SULFOTRANSFERASES , *METASTASIS , *TUMOR markers - Abstract
Metastasis is responsible for at least 90% of colon cancer (CC)‐related deaths. Lipid metabolism is a critical factor in cancer metastasis, yet the underlying mechanism requires further investigation. Herein, through the utilisation of single‐cell sequencing and proteomics, we identified sulfotransferase SULT2B1 as a novel metastatic tumour marker of CC, which was associated with poor prognosis. CC orthotopic model and in vitro assays showed that SULT2B1 promoted lipid metabolism and metastasis. Moreover, SULT2B1 directly interacted with SCD1 to facilitate lipid metabolism and promoted metastasis of CC cells. And the combined application of SCD1 inhibitor CAY with SULT2B1‐ konockout (KO) demonstrated a more robust inhibitory effect on lipid metabolism and metastasis of CC cells in comparison to sole application of SULT2B1‐KO. Notably, we revealed that lovastatin can block the SULT2B1‐induced promotion of lipid metabolism and distant metastasis in vivo. Further evidence showed that SMC1A transcriptionally upregulated the expression of SULT2B1. Our findings unveiled the critical role of SULT2B1 in CC metastasis and provided a new perspective for the treatment of CC patients with distant metastasis. [ABSTRACT FROM AUTHOR]
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- 2024
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5. Sulfotransferase SULT2B1 facilitates colon cancer metastasis by promoting SCD1‐mediated lipid metabolism
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Gang Che, Wankun Wang, Jiawei Wang, Cheng He, Jie Yin, Zhendong Chen, Chao He, Xujing Wang, Yan Yang, and Jian Liu
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colon cancer ,lipid metabolism ,metastasis ,SULT2B1 ,Medicine (General) ,R5-920 - Abstract
Abstract Metastasis is responsible for at least 90% of colon cancer (CC)‐related deaths. Lipid metabolism is a critical factor in cancer metastasis, yet the underlying mechanism requires further investigation. Herein, through the utilisation of single‐cell sequencing and proteomics, we identified sulfotransferase SULT2B1 as a novel metastatic tumour marker of CC, which was associated with poor prognosis. CC orthotopic model and in vitro assays showed that SULT2B1 promoted lipid metabolism and metastasis. Moreover, SULT2B1 directly interacted with SCD1 to facilitate lipid metabolism and promoted metastasis of CC cells. And the combined application of SCD1 inhibitor CAY with SULT2B1‐ konockout (KO) demonstrated a more robust inhibitory effect on lipid metabolism and metastasis of CC cells in comparison to sole application of SULT2B1‐KO. Notably, we revealed that lovastatin can block the SULT2B1‐induced promotion of lipid metabolism and distant metastasis in vivo. Further evidence showed that SMC1A transcriptionally upregulated the expression of SULT2B1. Our findings unveiled the critical role of SULT2B1 in CC metastasis and provided a new perspective for the treatment of CC patients with distant metastasis.
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- 2024
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6. Cholesterol sulfate limits neutrophil recruitment and gut inflammation during mucosal injury.
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Kenji Morino, Kazufumi Kunimura, Yuki Sugiura, Yoshihiro Izumi, Keisuke Matsubara, Sayaka Akiyoshi, Rae Maeda, Kenichiro Hirotani, Daiji Sakata, Seiya Mizuno, Satoru Takahashi, Takeshi Bamba, Takehito Uruno, and Yoshinori Fukui
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INFLAMMATORY bowel diseases ,INTESTINAL tumors ,NEUTROPHILS ,CHOLESTEROL ,SMALL intestine ,SODIUM sulfate ,PRESSURE ulcers - Abstract
During mucosal injury, intestinal immune cells play a crucial role in eliminating invading bacteria. However, as the excessive accumulation of immune cells promotes inflammation and delays tissue repair, it is essential to identify the mechanism that limits the infiltration of immune cells to the mucosal-luminal interface. Cholesterol sulfate (CS) is the lipid product of the sulfotransferase SULT2B1 and suppresses immune reactions by inhibiting DOCK2-mediated Rac activation. In this study, we aimed to elucidate the physiological role of CS in the intestinal tract. We found that, in the small intestine and colon, CS is predominantly produced in the epithelial cells close to the lumen. While dextran sodium sulfate (DSS)-induced colitis was exacerbated in Sult2b1-deficient mice with increased prevalence of neutrophils, the elimination of either neutrophils or intestinal bacteria in Sult2b1-deficient mice attenuated disease development. Similar results were obtained when the Dock2 was genetically deleted in Sult2b1-deficient mice. In addition, we also show that indomethacin-induced ulcer formation in the small intestine was exacerbated in Sult2b1-deficient mice and was ameliorated by CS administration. Thus, our results uncover that CS acts on inflammatory neutrophils, and prevents excessive gut inflammation by inhibiting the Rac activator DOCK2. The administration of CS may be a novel therapeutic strategy for inflammatory bowel disease and nonsteroidal anti-inflammatory drug-induced ulcers. [ABSTRACT FROM AUTHOR]
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- 2023
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7. The promoter hypermethylation of SULT2B1 accelerates esophagus tumorigenesis via downregulated PER1
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Zhuo Li, Meng‐Yan Li, Ling‐Ling Wang, Lei Li, Qing‐Yun Chen, Ying‐Hui Zhu, Yan Li, Yan‐ru Qin, and Xin‐Yuan Guan
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circadian rhythm ,ESCC ,hypermethylation ,proliferation ,SULT2B1 ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Abstract Background Esophageal cancer is currently the eighth most common tumor in the world and a leading cause of cancer death. SULT2B1 plays crucial roles in tumorigenesis. The purpose of this study is to explore the role of SULT2B1 in esophageal squamous cell carcinoma (ESCC). Methods The expression of SULT2B1 and its clinicopathological characteristics were evaluated in ESCC cohorts. Bisulfite genomic sequencing and methylation specific PCR were used to detect the promoter hypermethylation of the SULT2B1 gene. The effects of SULT2B1 on the biological characters of ESCC cells were identified on functional assays. Subcutaneous xenograft models revealed the role of SULT2B1 in vivo with tumor growth. RNA‐Seq analysis and qRT‐PCR were performed to recognize the targeted effect of SULT2B1 on PER1. Results SULT2B1 was not expressed or at a low level in most patients with ESCC or in ESCC cell lines, and this was accompanied by poor clinical prognosis. Furthermore, the downregulation of SULT2B1 occurred in promoter hypermethylation. According to the functional results, overexpression of SULT2B1 could inhibit tumoral proliferation in vitro and retard tumor growth in vivo, whereas SULT2B1 knockdown could accelerate ESCC progression. Mechanistically, SULT2B1 targeted PER1 at the mRNA level during post‐transcriptional regulation. Finally, PER1 was verified as a suppressor and poor‐prognosis factor in ESCC. Conclusions SULT2B1 loss is a consequence owing to its ability to promote hypermethylation. In addition, it serves as a suppressor and poor‐prognosis factor because of the post‐transcriptional regulation of PER1 in ESCC.
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- 2021
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8. Characterization of the mechanism of Scutellaria baicalensis on reversing radio-resistance in colorectal cancer
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Jinmei Huang, Ming Zhou, Huan Zhang, Yeying Fang, Gang Chen, Jiaying Wen, and LiMin Liu
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Colorectal cancer ,Radio-resistance ,Scutellaria baicalensis ,SULT2B1 ,Wogonin ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Scutellaria baicalensis (SB) has been shown to improve the therapeutic effects of colorectal cancer (CRC) and perform well for reversing radio-resistance in different cancers. However, its potential function and mechanism related to radio-resistance in CRC has not been explored. A radio-resistant human CRC cell line (HCT116R) was applied. A network pharmacological analysis was performed to reveal the potential mechanism of SB for reversing radio-resistance in CRC, and computational pathological analysis was applied to indicate the clinicopathological significance of the key targets. Then, our hypothesis was further verified by molecular docking. The network pharmacology analysis showed that wogonin is the key compound of SB for reversing the radio-resistance of CRC. A Kyoto Encyclopedia of Genes and Genomes analysis showed that the genes for SB that reverse radio-resistance in CRC are mainly involved in steroid hormone biosynthesis. An enrichment analysis pointed out that Sulfotransferase family 2B member 1 (SULT2B1) is a potentially vital gene. SULT2B1 was demonstrated as being highly expressed in CRC and upregulated in radio-resistant rectal tissues or cell lines. A CCK-8 and clone formation test showed that the viability and clone formation ability of HCT116R were significantly decreased by wogonin combined with radiotherapy, compared to radiotherapy alone. By contrast, flow cytometry revealed that the apoptosis of HCT116R was significantly increased when wogonin treatment combined with radiotherapy, compared with radiotherapy alone. Molecular docking verification indicated that SULT2B1 and wogonin have a good binding ability. Taken together, SULT2B1 may be the potential drug target in treating radio-resistant CRC. Wogonin may be the core compound of SB for reversing radio-resistance in CRC by targeting SULT2B1.
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- 2022
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9. The promoter hypermethylation of SULT2B1 accelerates esophagus tumorigenesis via downregulated PER1.
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Li, Zhuo, Li, Meng‐Yan, Wang, Ling‐Ling, Li, Lei, Chen, Qing‐Yun, Zhu, Ying‐Hui, Li, Yan, Qin, Yan‐ru, and Guan, Xin‐Yuan
- Abstract
Background Methods Results Conclusions Esophageal cancer is currently the eighth most common tumor in the world and a leading cause of cancer death. SULT2B1 plays crucial roles in tumorigenesis. The purpose of this study is to explore the role of SULT2B1 in esophageal squamous cell carcinoma (ESCC).The expression of SULT2B1 and its clinicopathological characteristics were evaluated in ESCC cohorts. Bisulfite genomic sequencing and methylation specific PCR were used to detect the promoter hypermethylation of the SULT2B1 gene. The effects of SULT2B1 on the biological characters of ESCC cells were identified on functional assays. Subcutaneous xenograft models revealed the role of SULT2B1 in vivo with tumor growth. RNA‐Seq analysis and qRT‐PCR were performed to recognize the targeted effect of SULT2B1 on PER1.SULT2B1 was not expressed or at a low level in most patients with ESCC or in ESCC cell lines, and this was accompanied by poor clinical prognosis. Furthermore, the downregulation of SULT2B1 occurred in promoter hypermethylation. According to the functional results, overexpression of SULT2B1 could inhibit tumoral proliferation in vitro and retard tumor growth in vivo, whereas SULT2B1 knockdown could accelerate ESCC progression. Mechanistically, SULT2B1 targeted PER1 at the mRNA level during post‐transcriptional regulation. Finally, PER1 was verified as a suppressor and poor‐prognosis factor in ESCC.SULT2B1 loss is a consequence owing to its ability to promote hypermethylation. In addition, it serves as a suppressor and poor‐prognosis factor because of the post‐transcriptional regulation of PER1 in ESCC. [ABSTRACT FROM AUTHOR]
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- 2021
- Full Text
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10. Downregulation of circDYNC1H1 exhibits inhibitor effect on cell proliferation and migration in hepatocellular carcinoma through miR‐140‐5p.
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Wang, Zheng‐Yang, Zhu, Zhu, Wang, Hua‐Fei, Qin, Bo, Liu, Jing, Yao, Xiao‐Han, Li, Wen‐Cai, and Chen, Kui‐Sheng
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CELL migration , *CELL proliferation , *HEPATOCELLULAR carcinoma , *CIRCULAR RNA , *MESSENGER RNA , *DOWNREGULATION - Abstract
Circular RNAs have been found to be aberrantly expressed in tumors and their significance in tumorigenesis has been focused on. The role of circDYNC1H1 in hepatocellular carcinoma (HCC) pathogenesis and its relationship with miR‐140‐5p were explored. The expression of circDYNC1H1, miR‐140‐5p, and SULT2B1 in HCC tissues and cells was measured, and Pearson's analysis was used to analyze their expression correlation. 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide and Transwell assays were performed to determine cell proliferation and migration. Binding between circDYNC1H1 and miR‐140‐5p was evaluated with RNA pull‐down assay. A luciferase reporter assay was conducted to assess the interaction between circDYNC1H1 and miR‐140‐5p and between miR‐140‐5p and SULT2B1. circDYNC1H1 was highly expressed in HCC tissues (n = 20), and it was negatively associated with the expression of miR‐140‐5p but positively correlated with SULT2B1 messenger RNA expression. circDYNC1H1 was upregulated in cell lines of HCC. Interference of circDYNC1H1 suppressed cell proliferation and migration of HCC. circDYNC1H1 acted as a sponge of miR‐140‐5p. miR‐140‐5p controlled SULT2B1 expression by targeting its 3′‐untranslated region. circDYNC1H1 enhanced SULT2B1 expression via sponging miR‐140‐5p. Downregulation of circDYNC1H1 disturbed cell proliferation and migration of HCC through miR‐140‐5p/SULT2B1 pathway. Silencing of circDYNC1H1 delayed tumor growth in HCC mouse model. Acting like a sponge of miR‐140‐5p, silenced circDYNC1H1 downregulated SULT2B1 to restrain HCC cell proliferation and migration, which is adverse to HCC growth and progression. [ABSTRACT FROM AUTHOR]
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- 2019
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11. Sult2b1 deficiency exacerbates ischemic stroke by promoting pro-inflammatory macrophage polarization in mice
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Yu Nie, Haojie Jin, Baohui Xu, Jihong Meng, Lixiang Xue, Feng Wang, Yafang Wang, Yan Wang, Xiaomu Tan, Heng Zhao, Yang Yao, and Cuixia Yang
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Male ,medicine.medical_specialty ,Macrophage ,Macrophage polarization ,Medicine (miscellaneous) ,Sult2b1 ,Cholesterol Sulfate ,Monocytes ,Brain Ischemia ,chemistry.chemical_compound ,Mice ,Immune system ,Neuroinflammation ,Internal medicine ,SULT2B1 ,Medicine ,Animals ,Humans ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Stroke ,Ischemic Stroke ,Inflammation ,Mice, Knockout ,business.industry ,Cholesterol ,Monocyte ,Macrophages ,Infarction, Middle Cerebral Artery ,Recovery of Function ,Macrophage Activation ,medicine.disease ,Mice, Inbred C57BL ,Disease Models, Animal ,medicine.anatomical_structure ,Endocrinology ,chemistry ,Neuroinflammatory Diseases ,Cholesterol Esters ,Microglia ,Sulfotransferases ,business ,Nicotinamide adenine dinucleotide phosphate ,Research Paper ,Signal Transduction - Abstract
Rationale: Stroke is a leading causes of human death worldwide. Ischemic damage induces the sterile neuroinflammation, which directly determines the recovery of patients. Lipids, a major component of the brain, significantly altered after stroke. Cholesterol sulfate, a naturally occurring analog of cholesterol, can directly regulate immune cell activation, indicating the possible involvement of cholesterol metabolites in neuroinflammation. Sulfotransferase family 2b member 1 (Sult2b1) is the key enzyme that catalyzes the synthesis of cholesterol sulfate. This study aimed to investigate the function of Sult2b1 and cholesterol sulfate in the neuroinflammation after ischemic stroke. Methods and Results: Sult2b1-/- and wild-type mice were subjected to transient middle cerebral artery occlusion. Our data showed that Sult2b1-/- mice had larger infarction and worse neurological scores. To determine whether immune cells were involved in the worsening stroke outcome in Sult2b1-/- mice, bone marrow transplantation, immune cell depletion, and adoptive monocyte transfer were performed. Combined with CyTOF and immunofluorescence techniques, we demonstrated that after stroke, the peripheral monocyte-derived macrophages were the dominant cell type promoting the pro-inflammatory status in Sult2b1-/-mice. Using primary bone marrow-derived macrophages, we showed that cholesterol sulfate could attenuate the pro-inflammatory polarization of macrophages under both normal and oxygen-glucose deprivation conditions by regulating the levels of nicotinamide adenine dinucleotide phosphate (NADPH), reactive oxygen species (ROS), and activating the AMP-activated protein kinase (AMPK) - cAMP responsive element-binding protein (CREB) signaling pathway. Conclusions: Sult2b1-/- promoted the polarization of macrophages into pro-inflammatory status. This trend could be attenuated by adding cholesterol sulfate, which promotes the polarization of macrophages into anti-inflammatory status by metabolic regulation. In this study, we established an inflammation-metabolism axis during the macrophage polarization after ischemic stroke.
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- 2021
12. Expression of SULT2A1, SULT2B1 and HSD3B1 in the porcine testis and epididymis.
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Zimmer, B., Tenbusch, L., DKlymiuk, M. C., Dezhkam, Y ., and Schuler, G .
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GENE expression , *SULFOTRANSFERASES , *DEHYDROEPIANDROSTERONE , *POLYMERASE chain reaction , *IMMUNOHISTOCHEMISTRY , *EPIDIDYMIS - Abstract
In the porcine testis, in addition to estrogen sulfates, the formation of numerous sulfonated neutral hydroxysteroids has been observed. However, their functions and the underlying synthetic pathways are still widely unclear. To obtain further information on their formation in postpubertal boars, the expression of sulfotransferases considered relevant for neutral hydroxysteroids (SULT2A1, SULT2B1) was investigated in the testis and defined segments of the epididymis applying real-time RT-qPCR, Western blot and immunohistochemistry (IHC). Sulfotransferase activities were assessed in tissue homogenates or cytosolic preparations applying dehydroepiandrosterone and pregnenolone as substrates. A high SULT2A1 expression was confirmed in the testis and localized in Leydig cells by IHC. In the epididymis, SULT2A1 expression was virtually confined to the body. SULT2B1 expression was absent or low in the testis but increased significantly along the epididymis. Immunohistochemical observations indicate that both enzymes are secreted into the ductal lumen via an apocrine mechanism. The results from the characterization of expression patterns and activity measurements suggest that SULT2A1 is the prevailing enzyme for the sulfonation of hydroxysteroids in the testis, whereas SULT2B1 may catalyze the formation of sterol sulfates in the epididymis. In order to obtain information on the overall steroidogenic capacity of the porcine epididymis, the expression of important steroidogenic enzymes (CYP11A1, CYP17A1, CYP19, HSD3B1, HSD17B3, SRD5A2) was monitored in the defined epididymal segments applying real-time RT-qPCR. Surprisingly, in addition to a high expression of SRD5A2 in the epididymal head, a substantial expression of HSD3B1 was detected, which increased along the organ. [ABSTRACT FROM AUTHOR]
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- 2018
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13. Association study of genetic variants in estrogen metabolic pathway genes and colorectal cancer risk and survival.
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Li, Shuwei, Xie, Lisheng, Du, Mulong, Xu, Kaili, Zhu, Lingjun, Chu, Haiyan, Chen, Jinfei, Wang, Meilin, Zhang, Zhengdong, and Gu, Dongying
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COLON cancer , *ESTROGEN , *SINGLE nucleotide polymorphisms , *LOGISTIC regression analysis , *DISEASE progression - Abstract
Although studies have investigated the association of genetic variants and the abnormal expression of estrogen-related genes with colorectal cancer risk, the evidence remains inconsistent. We clarified the relationship of genetic variants in estrogen metabolic pathway genes with colorectal cancer risk and survival. A case-control study was performed to assess the association of single-nucleotide polymorphisms (SNPs) in ten candidate genes with colorectal cancer risk in a Chinese population. A logistic regression model and Cox regression model were used to calculate SNP effects on colorectal cancer susceptibility and survival, respectively. Expression quantitative trait loci (eQTL) analysis was conducted using the Genotype-Tissue Expression (GTEx) project dataset. The sequence kernel association test (SKAT) was used to perform gene-set analysis. Colorectal cancer risk and rs3760806 in SULT2B1 were significantly associated in both genders [male: OR = 1.38 (1.15-1.66); female: OR = 1.38 (1.13-1.68)]. Two SNPs in SULT1E1 were related to progression-free survival (PFS) [rs1238574: HR = 1.24 (1.02-1.50), P = 2.79 × 10−2; rs3822172: HR = 1.30 (1.07-1.57), P = 8.44 × 10−3] and overall survival (OS) [rs1238574: HR = 1.51 (1.16-1.97), P = 2.30 × 10−3; rs3822172: HR = 1.53 (1.67-2.00), P = 2.03 × 10−3]. Moreover, rs3760806 was an eQTL for SULT2B1 in colon samples (transverse: P = 3.6 × 10−3; sigmoid: P = 1.0 × 10−3). SULT2B1 expression was significantly higher in colorectal tumor tissues than in normal tissues in the Cancer Genome Atlas (TCGA) database (P < 1.0 × 10−4). Our results indicated that SNPs in estrogen metabolic pathway genes confer colorectal cancer susceptibility and survival. [ABSTRACT FROM AUTHOR]
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- 2018
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14. Sulfation of vitamin D3-related compounds-identification and characterization of the responsible human cytosolic sulfotransferases.
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Kurogi, Katsuhisa, Sakakibara, Yoichi, Suiko, Masahito, and Liu, Ming-Cheh
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SULFATION , *ADDITION reactions , *PHYSIOLOGICAL effects of vitamin D , *PHYSIOLOGICAL effects of vitamins , *SULFOTRANSFERASES - Abstract
While 25-hydroxyvitamin D3 3- O-sulfate is known to be present in circulation, how it is generated in the body remains unclear. This study aimed to investigate its sulfation in major human organs and to unveil the responsible cytosolic sulfotransferases ( SULTs). Of the vitamin D3-related compounds tested, 25-hydroxyvitamin D3 and 7-dehydrocholesterol are preferentially sulfated by human organ cytosols. Among the 13 human SULTs, SULT2A1 shows sulfating activity toward all vitamin D3-related compounds, whereas SULT1A1 and SULT2B1a/ SULT2B1b show sulfating activity exclusively for, respectively, calcitriol and 7-dehydrocholesterol. These findings suggest that the metabolic pathway leading to the formation of 25-hydroxyvitamin D3 3- O-sulfate may be mediated by the sulfation of 25-hydroxyvitamin D3 or by the conversion of 7-dehydrocholesterol-3- O-sulfate in the skin. [ABSTRACT FROM AUTHOR]
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- 2017
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15. Mutations in SULT2B1 Cause Autosomal-Recessive Congenital Ichthyosis in Humans.
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Heinz, Lisa, Kim, Gwang-Jin, Marrakchi, Slaheddine, Christiansen, Julie, Turki, Hamida, Rauschendorf, Marc-Alexander, Lathrop, Mark, Hausser, Ingrid, Zimmer, Andreas D., and Fischer, Judith
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CONGENITAL ichthyosiform erythroderma , *GENETIC mutation , *SKIN physiology , *KERATINOCYTES , *FIBROBLASTS - Abstract
Ichthyoses are a clinically and genetically heterogeneous group of genodermatoses associated with abnormal scaling of the skin over the whole body. Mutations in nine genes are known to cause non-syndromic forms of autosomal-recessive congenital ichthyosis (ARCI). However, not all genetic causes for ARCI have been discovered to date. Using whole-exome sequencing (WES) and multigene panel screening, we identified 6 ARCI-affected individuals from three unrelated families with mutations in Sulfotransferase family 2B member 1 ( SULT2B1 ), showing their causative association with ARCI. Cytosolic sulfotransferases form a large family of enzymes that are involved in the synthesis and metabolism of several steroids in humans. We identified four distinct mutations including missense, nonsense, and splice site mutations. We demonstrated the loss of SULT2B1 expression at RNA and protein levels in keratinocytes from individuals with ARCI by functional analyses. Furthermore, we succeeded in reconstructing the morphologic skin alterations in a 3D organotypic tissue culture model with SULT2B1 -deficient keratinocytes and fibroblasts. By thin layer chromatography (TLC) of extracts from these organotypic cultures, we could show the absence of cholesterol sulfate, the metabolite of SULT2B1, and an increased level of cholesterol, indicating a disturbed cholesterol metabolism of the skin upon loss-of-function mutation in SULT2B1 . In conclusion, our study reveals an essential role for SULT2B1 in the proper development of healthy human skin. Mutation in SULT2B1 leads to an ARCI phenotype via increased proliferation of human keratinocytes, thickening of epithelial layers, and altered epidermal cholesterol metabolism. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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16. Cholesterol sulfate limits neutrophil recruitment and gut inflammation during mucosal injury.
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Morino K, Kunimura K, Sugiura Y, Izumi Y, Matsubara K, Akiyoshi S, Maeda R, Hirotani K, Sakata D, Mizuno S, Takahashi S, Bamba T, Uruno T, and Fukui Y
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- Animals, Mice, Neutrophil Infiltration, Guanine Nucleotide Exchange Factors, GTPase-Activating Proteins, Inflammation, Colitis
- Abstract
During mucosal injury, intestinal immune cells play a crucial role in eliminating invading bacteria. However, as the excessive accumulation of immune cells promotes inflammation and delays tissue repair, it is essential to identify the mechanism that limits the infiltration of immune cells to the mucosal-luminal interface. Cholesterol sulfate (CS) is the lipid product of the sulfotransferase SULT2B1 and suppresses immune reactions by inhibiting DOCK2-mediated Rac activation. In this study, we aimed to elucidate the physiological role of CS in the intestinal tract. We found that, in the small intestine and colon, CS is predominantly produced in the epithelial cells close to the lumen. While dextran sodium sulfate (DSS)-induced colitis was exacerbated in Sult2b1 -deficient mice with increased prevalence of neutrophils, the elimination of either neutrophils or intestinal bacteria in Sult2b1 -deficient mice attenuated disease development. Similar results were obtained when the Dock2 was genetically deleted in Sult2b1 -deficient mice. In addition, we also show that indomethacin-induced ulcer formation in the small intestine was exacerbated in Sult2b1 -deficient mice and was ameliorated by CS administration. Thus, our results uncover that CS acts on inflammatory neutrophils, and prevents excessive gut inflammation by inhibiting the Rac activator DOCK2. The administration of CS may be a novel therapeutic strategy for inflammatory bowel disease and non-steroidal anti-inflammatory drug-induced ulcers., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Morino, Kunimura, Sugiura, Izumi, Matsubara, Akiyoshi, Maeda, Hirotani, Sakata, Mizuno, Takahashi, Bamba, Uruno and Fukui.)
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- 2023
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17. An in vitro investigation of endocrine disrupting potentials of ten bisphenol analogues
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Yao Wang, Sang-Woo Lee, Kyungho Choi, Xiaoshan Liu, Juntong Lin, Mingwei Sun, and Langjing Deng
- Subjects
endocrine system ,Sulfotransferase ,Bisphenol ,Clinical Biochemistry ,030209 endocrinology & metabolism ,Pharmacology ,Endocrine Disruptors ,Biochemistry ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Phenols ,SULT2B1 ,Potency ,Bioassay ,Luciferase ,Benzhydryl Compounds ,Molecular Biology ,Reporter gene ,urogenital system ,Chemistry ,Organic Chemistry ,030220 oncology & carcinogenesis ,Toxicity ,Biological Assay ,hormones, hormone substitutes, and hormone antagonists - Abstract
The endocrine disruption potency of BPA was reported elsewhere, but the mechanisms of its analogues have not been fully resolved. In this study, endocrine disruption potentials of nine alternative bisphenol analogues, namely 2,2-bis(4-hydroxyphenyl)butane (BPB), 2,2-Bis(4-hydroxy-3-methylphenyl)propane (BPC), 4,4′-dihydroxydiphenylmethane (BPF), 4,4′-(1,3-Phenylene diisopropylidene)bisphenol (BPM), 4,4′-(1,4-phenylenediisopropylidene)bisphenol (BPP), 4,4′- sulfonyldiphenol (BPS), 4,4′ cyclohexylidenebisphenol (BPZ), 4,4′ (hexafluoroisopropylidene)-diphenol (BPAF) and 4,4′-(1-phenylethylidene)bisphenol (BPAP), plus 2,2-bis(4-hydroxyphenyl)propane (BPA) were investigated by H295R cell and MVLN cell bioassays. In the H295R cell assay, the endpoints included hormone production and key genes for steroidogenesis (CYP11A, CYP17, CYP19 and 3βHSD2) or metabolism sulfotransferase (SULT1A1, SULT2A1 and SULT2B1) at the molecular level. The results indicated that except for BPP or BPAF, the eight other bisphenols significantly increased the E2/T ratio. In addition, BPB, BPF and BPS significantly up-regulate CYP19 gene expression, and only BPB significantly reduced sulfotransferase gene expression. In the MVLN luciferase gene reporter assay, seven bisphenols induced luciferase activity alone, and are 104 to 108-fold less potent than E2. Their nuclear ERα binding activity is in the order of BPAF > BPZ > BPP > BPB > BPA > BPF > BPS. In summary, all nine tested bisphenols showed endocrine toxicity through different mechanisms. Some had similar potency as BPA, but some had even higher potency. Further research is necessary to evaluate the toxicity of these potential BPA substitutes.
- Published
- 2020
18. Characterization of the mechanism of Scutellaria baicalensis on reversing radio-resistance in colorectal cancer.
- Author
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Huang J, Zhou M, Zhang H, Fang Y, Chen G, Wen J, and Liu L
- Abstract
Scutellaria baicalensis (SB) has been shown to improve the therapeutic effects of colorectal cancer (CRC) and perform well for reversing radio-resistance in different cancers. However, its potential function and mechanism related to radio-resistance in CRC has not been explored. A radio-resistant human CRC cell line (HCT116R) was applied. A network pharmacological analysis was performed to reveal the potential mechanism of SB for reversing radio-resistance in CRC, and computational pathological analysis was applied to indicate the clinicopathological significance of the key targets. Then, our hypothesis was further verified by molecular docking. The network pharmacology analysis showed that wogonin is the key compound of SB for reversing the radio-resistance of CRC. A Kyoto Encyclopedia of Genes and Genomes analysis showed that the genes for SB that reverse radio-resistance in CRC are mainly involved in steroid hormone biosynthesis. An enrichment analysis pointed out that Sulfotransferase family 2B member 1 (SULT2B1) is a potentially vital gene. SULT2B1 was demonstrated as being highly expressed in CRC and upregulated in radio-resistant rectal tissues or cell lines. A CCK-8 and clone formation test showed that the viability and clone formation ability of HCT116R were significantly decreased by wogonin combined with radiotherapy, compared to radiotherapy alone. By contrast, flow cytometry revealed that the apoptosis of HCT116R was significantly increased when wogonin treatment combined with radiotherapy, compared with radiotherapy alone. Molecular docking verification indicated that SULT2B1 and wogonin have a good binding ability. Taken together, SULT2B1 may be the potential drug target in treating radio-resistant CRC. Wogonin may be the core compound of SB for reversing radio-resistance in CRC by targeting SULT2B1., (Copyright © 2022. Published by Elsevier Inc.)
- Published
- 2022
- Full Text
- View/download PDF
19. SULFATION PATHWAYS: Expression of SULT2A1, SULT2B1 and HSD3B1 in the porcine testis and epididymis
- Author
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L Tenbusch, Y Dezhkam, Gerhard Schuler, B Zimmer, and MC Klymiuk
- Subjects
0301 basic medicine ,Sulfotransferase ,030219 obstetrics & reproductive medicine ,Chemistry ,Cholesterol side-chain cleavage enzyme ,Epididymis ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Endocrinology ,medicine.anatomical_structure ,CYP17A1 ,SULT2B1 ,HSD3B1 ,medicine ,Pregnenolone ,Molecular Biology ,Hydroxysteroids ,medicine.drug - Abstract
In the porcine testis, in addition to estrogen sulfates, the formation of numerous sulfonated neutral hydroxysteroids has been observed. However, their functions and the underlying synthetic pathways are still widely unclear. To obtain further information on their formation in postpubertal boars, the expression of sulfotransferases considered relevant for neutral hydroxysteroids (SULT2A1, SULT2B1) was investigated in the testis and defined segments of the epididymis applying real-time RT-qPCR, Western blot and immunohistochemistry (IHC). Sulfotransferase activities were assessed in tissue homogenates or cytosolic preparations applying dehydroepiandrosterone and pregnenolone as substrates. A highSULT2A1expression was confirmed in the testis and localized in Leydig cells by IHC. In the epididymis,SULT2A1expression was virtually confined to the body.SULT2B1expression was absent or low in the testis but increased significantly along the epididymis. Immunohistochemical observations indicate that both enzymes are secreted into the ductal lumen via an apocrine mechanism. The results from the characterization of expression patterns and activity measurements suggest that SULT2A1 is the prevailing enzyme for the sulfonation of hydroxysteroids in the testis, whereas SULT2B1 may catalyze the formation of sterol sulfates in the epididymis. In order to obtain information on the overall steroidogenic capacity of the porcine epididymis, the expression of important steroidogenic enzymes (CYP11A1, CYP17A1, CYP19, HSD3B1, HSD17B3, SRD5A2) was monitored in the defined epididymal segments applying real-time RT-qPCR. Surprisingly, in addition to a high expression ofSRD5A2in the epididymal head, a substantial expression ofHSD3B1was detected, which increased along the organ.
- Published
- 2018
20. Hydroxysteroid sulfotransferase 2B1 affects gastric epithelial function and carcinogenesis induced by a carcinogenic agent
- Author
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Ziyan Zhuang, Mingjie Yang, Wenting Hong, Fenghua Guo, Dongke Xu, Baolin Niu, Xiaobo Li, and Qianming Bai
- Subjects
0301 basic medicine ,Gastric epithelial cell ,Carcinogenesis ,Endocrinology, Diabetes and Metabolism ,Clinical Biochemistry ,medicine.disease_cause ,Mice ,Phosphatidylinositol 3-Kinases ,0302 clinical medicine ,Endocrinology ,RNA, Small Interfering ,lcsh:RC620-627 ,Gene Editing ,Mice, Knockout ,Chemistry ,digestive, oral, and skin physiology ,Gene Expression Regulation, Neoplastic ,lcsh:Nutritional diseases. Deficiency diseases ,Cholesterol ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Sulfotransferases ,PI3K/AKT signaling ,Signal Transduction ,Hydroxysteroid Sulfotransferase 2B1 (SULT2B1) ,Oxysterol ,03 medical and health sciences ,Stomach Neoplasms ,Cell Line, Tumor ,SULT2B1 ,medicine ,Animals ,Humans ,Protein kinase B ,Gastric carcinogenesis ,Carcinogen ,PI3K/AKT/mTOR pathway ,Cell Proliferation ,Base Sequence ,Akt/PKB signaling pathway ,Research ,Biochemistry (medical) ,Survival Analysis ,digestive system diseases ,Hydroxycholesterols ,Epithelium ,030104 developmental biology ,Gastric Mucosa ,Cancer research ,CRISPR-Cas Systems ,Proto-Oncogene Proteins c-akt ,Methylcholanthrene - Abstract
Background A healthy gastric mucosal epithelium exhibits tumor-suppressive properties. Gastric epithelial cell dysfunction contributes to gastric cancer development. Oxysterols provided from food or cholesterol oxidation in the gastric epithelium may be further sulfated by hydroxysteroid sulfotransferase 2B1 (SULT2B1), which is highly abundant in the gastric epithelium. However, the effects of SULT2B1 on gastric epithelial function and gastric carcinogenesis are unclear. Methods A mouse gastric tumor model was established using carcinogenic agent 3-methylcholanthrene (3-MCA). A SULT2B1 deletion (SULT2B1−/−) human gastric epithelial line GES-1 was constructed by CRISPR/CAS9 genome editing system. Results The gastric tumor incidence was higher in the SULT2B1−/− mice than in the wild-type (WT) mice. In gastric epithelial cells, adenovirus-mediated SULT2B1b overexpression reduced the levels of oxysterols, such as 24(R/S),25-epoxycholesterol (24(R/S),25-EC) and 27-hydroxycholesterol (27HC). This condition also increased PI3K/AKT signaling to promote gastric epithelial cell proliferation, epithelization, and epithelial development. However, SULT2B1 deletion or SULT2B1 knockdown suppressed PI3K/AKT signaling, epithelial cell epithelization, and wound healing and induced gastric epithelial cell malignant transition upon 3-MCA induction. Conclusions The abundant SULT2B1 expression in normal gastric epithelium might maintain epithelial function via the PI3K/AKT signaling pathway and suppress gastric carcinogenesis induced by a carcinogenic agent.
- Published
- 2019
21. Sulfation of vitamin D3-related compounds-identification and characterization of the responsible human cytosolic sulfotransferases
- Author
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Ming-Cheh Liu, Masahito Suiko, Yoichi Sakakibara, and Katsuhisa Kurogi
- Subjects
0301 basic medicine ,Vitamin ,Calcitriol ,Chemistry ,Biophysics ,Cell Biology ,Body remains ,Biochemistry ,03 medical and health sciences ,Metabolic pathway ,Cytosol ,7-Dehydrocholesterol ,chemistry.chemical_compound ,030104 developmental biology ,0302 clinical medicine ,Sulfation ,Structural Biology ,030220 oncology & carcinogenesis ,SULT2B1 ,polycyclic compounds ,Genetics ,medicine ,Molecular Biology ,medicine.drug - Abstract
While 25-hydroxyvitamin D3 3-O-sulfate is known to be present in circulation, how it is generated in the body remains unclear. This study aimed to investigate its sulfation in major human organs and to unveil the responsible cytosolic sulfotransferases (SULTs). Of the vitamin D3 -related compounds tested, 25-hydroxyvitamin D3 and 7-dehydrocholesterol are preferentially sulfated by human organ cytosols. Among the 13 human SULTs, SULT2A1 shows sulfating activity toward all vitamin D3 -related compounds, whereas SULT1A1 and SULT2B1a/SULT2B1b show sulfating activity exclusively for, respectively, calcitriol and 7-dehydrocholesterol. These findings suggest that the metabolic pathway leading to the formation of 25-hydroxyvitamin D3 3-O-sulfate may be mediated by the sulfation of 25-hydroxyvitamin D3 or by the conversion of 7-dehydrocholesterol-3-O-sulfate in the skin.
- Published
- 2017
22. Sult2b1 deficiency exacerbates ischemic stroke by promoting pro-inflammatory macrophage polarization in mice.
- Author
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Wang Y, Jin H, Wang Y, Yao Y, Yang C, Meng J, Tan X, Nie Y, Xue L, Xu B, Zhao H, and Wang F
- Subjects
- Animals, Brain Ischemia metabolism, Cholesterol Esters genetics, Disease Models, Animal, Humans, Infarction, Middle Cerebral Artery, Inflammation metabolism, Ischemic Stroke metabolism, Macrophage Activation drug effects, Macrophages metabolism, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Microglia metabolism, Monocytes drug effects, Neuroinflammatory Diseases physiopathology, Recovery of Function drug effects, Signal Transduction drug effects, Stroke metabolism, Sulfotransferases genetics, Sulfotransferases metabolism, Cholesterol Esters metabolism, Ischemic Stroke genetics, Sulfotransferases deficiency
- Abstract
Rationale: Stroke is a leading causes of human death worldwide. Ischemic damage induces the sterile neuroinflammation, which directly determines the recovery of patients. Lipids, a major component of the brain, significantly altered after stroke. Cholesterol sulfate, a naturally occurring analog of cholesterol, can directly regulate immune cell activation, indicating the possible involvement of cholesterol metabolites in neuroinflammation. Sulfotransferase family 2b member 1 (Sult2b1) is the key enzyme that catalyzes the synthesis of cholesterol sulfate. This study aimed to investigate the function of Sult2b1 and cholesterol sulfate in the neuroinflammation after ischemic stroke. Methods and Results : Sult2b1
-/- and wild-type mice were subjected to transient middle cerebral artery occlusion. Our data showed that Sult2b1-/- mice had larger infarction and worse neurological scores. To determine whether immune cells were involved in the worsening stroke outcome in Sult2b1-/- mice, bone marrow transplantation, immune cell depletion, and adoptive monocyte transfer were performed. Combined with CyTOF and immunofluorescence techniques, we demonstrated that after stroke, the peripheral monocyte-derived macrophages were the dominant cell type promoting the pro-inflammatory status in Sult2b1-/- mice. Using primary bone marrow-derived macrophages, we showed that cholesterol sulfate could attenuate the pro-inflammatory polarization of macrophages under both normal and oxygen-glucose deprivation conditions by regulating the levels of nicotinamide adenine dinucleotide phosphate (NADPH), reactive oxygen species (ROS), and activating the AMP-activated protein kinase (AMPK) - cAMP responsive element-binding protein (CREB) signaling pathway. Conclusions: Sult2b1-/- promoted the polarization of macrophages into pro-inflammatory status. This trend could be attenuated by adding cholesterol sulfate, which promotes the polarization of macrophages into anti-inflammatory status by metabolic regulation. In this study, we established an inflammation-metabolism axis during the macrophage polarization after ischemic stroke., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)- Published
- 2021
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23. Cholesterol sulfate is a DOCK2 inhibitor that mediates tissue-specific immune evasion in the eye
- Author
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Miho Ushijima, Takaaki Tatsuguchi, Takehito Uruno, Kazuhiko Yamamura, Makoto Suematsu, Yuki Sugiura, Chiemi Mishima-Tsumagari, Yoshinori Fukui, Mayuki Watanabe, Yuko Hattori, Mutsuko Kukimoto-Niino, and Tetsuya Sakurai
- Subjects
0301 basic medicine ,Leukocyte migration ,Serine Proteinase Inhibitors ,Inflammation ,Eye ,Biochemistry ,Mice ,03 medical and health sciences ,Harderian gland ,Sulfation ,Immune system ,SULT2B1 ,medicine ,Animals ,Guanine Nucleotide Exchange Factors ,Photosensitivity Disorders ,Molecular Biology ,Immune Evasion ,Keratitis ,Mice, Knockout ,biology ,Chemistry ,Dock2 ,GTPase-Activating Proteins ,Cell Biology ,Cell biology ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,biology.protein ,Cholesterol Esters ,Guanine nucleotide exchange factor ,Sulfotransferases ,medicine.symptom - Abstract
Although immune responses are essential to protect the body from infection, they can also harm tissues. Certain tissues and organs, including the eye, constitute specialized microenvironments that locally inhibit immune reactivity. Dedicator of cytokinesis protein 2 (DOCK2) is a Rac-specific guanine nucleotide exchange factor (GEF) that is predominantly found in hematopoietic cells. DOCK2 plays a key role in immune surveillance because it is essential for the activation and migration of leukocytes. DOCK2 mutations cause severe immunodeficiency in humans. We found that DOCK2-mediated Rac activation and leukocyte migration were effectively inhibited by cholesterol sulfate (CS), but not by cholesterol or other sulfated steroids. CS bound to the catalytic domain of DOCK2 and suppressed its GEF activity. Mass spectrometric quantification revealed that CS was most abundantly produced in the Harderian gland, which provides the lipids that form the oily layer of the tear film. Sulfation of cholesterol is mediated by the sulfotransferases SULT2B1b and, to a lesser extent, SULT2B1a, which are produced from the same gene through alternative splicing. By genetically inactivating Sult2b1, we showed that the lack of CS in mice augmented ultraviolet- and antigen-induced ocular surface inflammation, which was suppressed by administration of eye drops containing CS. Thus, CS is a naturally occurring DOCK2 inhibitor and contributes to the generation of the immunosuppressive microenvironment in the eye.
- Published
- 2018
24. Effects of human sulfotransferases on the cytotoxicity of 12-hydroxynevirapine
- Author
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Gonçalo Gamboa da Costa, Lucie Loukotková, Priyanka Chitranshi, Frederick A. Beland, and Jia-Long Fang
- Subjects
0301 basic medicine ,Male ,Nevirapine ,Cell Survival ,030106 microbiology ,Biochemistry ,Rats, Sprague-Dawley ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,SULT2B1 ,medicine ,Cytotoxic T cell ,Animals ,Humans ,Cytotoxicity ,Pharmacology ,Reverse-transcriptase inhibitor ,Dose-Response Relationship, Drug ,Chemistry ,Cytotoxins ,Glutathione ,Molecular biology ,Rats ,Mice, Inbred C57BL ,030104 developmental biology ,HEK293 Cells ,Cell culture ,Sulfotransferases ,Cysteine ,medicine.drug - Abstract
Nevirapine, a non-nucleoside reverse transcriptase inhibitor used for the treatment of AIDS, can cause serious skin rashes and hepatotoxicity. Previous studies have indicated that the benzylic sulfate 12-sulfoxynevirapine, the formation of which is catalyzed by human sulfotransferases (SULTs), may play a causative role in these toxicities. To characterize better the role of 12-sulfoxynevirapine in nevirapine-induced cytotoxicity, the ability of 12 expressed human SULT isoforms to conjugate 12-hydroxynevirapine was assessed. Of the 12 human SULTs, no detectable 12-sulfoxynevirapine was observed with SULT1A3, SULT1C2, SULT1C3, SULT2B1, SULT4A1, or SULT6B1. As determined by the Vmax/Km ratio, SULT2A1 had the highest overall 12-hydoxynevirapine sulfonation activity; lower activities were observed with SULT1A1, SULT1A2, SULT1B1, SULT1C4, and SULT1E1. Incubation of 12-sulfoxynevirapine with glutathione and cysteine led to adduct formation; lower yields were obtained with deoxynucleosides. 12-Hydroxynevirapine was more cytotoxic than nevirapine to TK6, TK6/SULT vector, and TK6/SULT2A1 cells. With nevirapine, there was no difference in cytotoxicity among the three cell lines, whereas with 12-hydroxynevirapine, TK6/SULT2A1 cells were more resistant than TK6 and TK6/SULT vector cells. Co-incubation of 12-hydroxynevirapine with the competitive SULT2A1 substrate dehydroepiandrosterone decreased the level of 12-sulfoxynevirapine and increased the cytotoxicity in TK6/SULT2A1 cells. These data demonstrate that although 12-sulfoxynevirapine reacts with nucleophiles to form adducts, sulfonation of 12-hydroxynevirapine decreases the cytotoxicity of 12-hydroxynevirapine in TK6 cells.
- Published
- 2018
25. Effect of SULT2B1 genetic polymorphisms on the sulfation of dehydroepiandrosterone and pregnenolone by SULT2B1b allozymes
- Author
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Katsuhisa Kurogi, Mohammed I. Rasool, Masahito Suiko, Amal A. El Daibani, Ahsan F. Bairam, Yoichi Sakakibara, Ming-Cheh Liu, Maryam S. Abunnaja, and Fatemah A. Alherz
- Subjects
0301 basic medicine ,endocrine system ,medicine.medical_specialty ,medicine.medical_treatment ,Dehydroepiandrosterone ,030209 endocrinology & metabolism ,Polymorphism, Single Nucleotide ,Biochemistry ,Article ,Steroid ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Sulfation ,Internal medicine ,SULT2B1 ,polycyclic compounds ,medicine ,Humans ,skin and connective tissue diseases ,Molecular Biology ,chemistry.chemical_classification ,Isoenzymes ,030104 developmental biology ,Enzyme ,chemistry ,Pregnenolone ,Sulfotransferases ,Hydroxysteroids ,hormones, hormone substitutes, and hormone antagonists ,medicine.drug ,Hormone - Abstract
Pregnenolone and dehydroepiandrosterone (DHEA) are hydroxysteroids that serve as biosynthetic precursors for steroid hormones in human body. SULT2B1b has been reported to be critically involved in the sulfation of pregnenolone and DHEA, particularly in the sex steroid-responsive tissues. The current study was designed to investigate the impact of the genetic polymorphisms of SULT2B1 on the sulfation of DHEA and pregnenolone by SULT2B1b allozymes. Ten SULT2B1b allozymes previously prepared were shown to exhibit differential sulfating activities toward DHEA and pregnenolone in comparison to the wild-type enzyme. Kinetic studies revealed further significant changes in their substrate-binding affinity and catalytic activity toward DHEA and pregnenolone. Taken together, these results indicated clearly a profound effect of SULT2B1 genetic polymorphisms on the sulfating activity of SULT2B1b allozymes toward DHEA and pregnenolone, which may have implications in inter-individual variations in the homeostasis of these two important steroid precursors.
- Published
- 2019
26. Mouse cytosolic sulfotransferase SULT2B1b interacts with cytoskeletal proteins via a proline/serine-rich C-terminus
- Author
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Yosuke Kamemoto, Ming-Cheh Liu, Katsuhisa Kurogi, Saki Takahashi, Masahito Suiko, Shin Yasuda, and Yoichi Sakakibara
- Subjects
chemistry.chemical_classification ,Sulfotransferase ,C-terminus ,macromolecular substances ,Cell Biology ,Biology ,Biochemistry ,Amino acid ,Serine ,Sulfation ,chemistry ,SULT2B1 ,Cytoskeleton ,Molecular Biology ,Actin - Abstract
Cytosolic sulfotransferase (SULT) SULT2B1b had previously been characterized as a cholesterol sulfotransferase. Like human SULT2B1, mouse SULT2B1b contains a unique, 31 amino acid C-terminal sequence with a proline/serine-rich region, which is not found in members of other SULT families. To gain insight into the functional relevance of this proline/serine-rich region, we constructed a truncated mouse SULT2B1b lacking the 31 C-terminal amino acids, and compared it with the wild-type enzyme. Enzymatic characterization indicated that the catalytic activity was not significantly affected by the absence of those C-terminal residues. Glutathione S-transferase pulldown assays showed that several proteins interacted with mouse SULT2B1b specifically through this C-terminal proline/serine-rich region. Peptide mass fingerprinting revealed that of the five SULT2B1b-binding proteins analyzed, three were cytoskeletal proteins and two were cytoskeleton-binding molecular chaperones. Furthermore, wild-type mouse SULT2B1b, but not the truncated enzyme, was associated with the cytoskeleton in experiments with a cytoskeleton-stabilizing buffer. Collectively, these results suggested that the unique, extended proline/serine-rich C-terminus of mouse SULT2B1b is important for its interaction with cytoskeletal proteins. Such an interaction may allow the enzyme to move along microfilaments such as actin filaments, and catalyze the sulfation of hydroxysteroids, such as cholesterol and pregnenolone, at specific intracellular locations. Structured digital abstract • MINT-7975854: Sult2B1b (uniprotkb:O35400) physically interacts (MI:0914) with Myosin-Ic (uniprotkb:Q9WTI7), Alpha-actinin-1 (uniprotkb:Q7TPR4), Alpha-actinin-4 (uniprotkb:P57780), HSP 90-beta (uniprotkb:P11499), Hsc70, (uniprotkb:P63017), Beta-actin (uniprotkb:P60710) and Gamma-actin (uniprotkb:P63260) by pull down (MI:0096)
- Published
- 2010
27. Effect of sex steroids on expression of sulfotransferase 2B1 immunoreactive protein in primary cultured porcine hepatocytes
- Author
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M. Moe, N. Panella-Riera, M. A. Oliver, Olena Doran, E. Grindflek, and J. D. Wood
- Subjects
Sulfotransferase ,medicine.medical_specialty ,General Veterinary ,Boar taint ,medicine.medical_treatment ,Androstenone ,Adipose tissue ,Metabolism ,Biology ,Steroid ,chemistry.chemical_compound ,Endocrinology ,chemistry ,SULT2B1 ,Internal medicine ,medicine ,Animal Science and Zoology ,Testosterone - Abstract
An excessive accumulation of androstenone (5α-androst-16-en-3-one) in pig adipose tissue is one of the two major contributors to the phenomenon of boar taint. High levels of adipose tissue androstenone have been related to a low rate of hepatic androstenone metabolism, which includes two stages: oxidative and conjugative. Sulfotransferases (SULTs), alongside with other specific enzymes, play the key role in the conjugative stage of androstenone metabolism. The present study investigated the mechanism regulating expression of sulfotransferase 2B1 (SULT2B1) immunoreactive protein using primary cultured pig hepatocytes as a model system. A specific objective was to determine whether the expression of pig hepatic SULT2B1 is regulated by the sex steroids; androstenone, testosterone and estrone sulphate. The study was performed on entire male pigs of a Large White (40%) × Landrace (40%) × Duroc (20%) cross-breed, average carcass weight 72.2 kg. The study shows that SULT2B1 immunoreactive protein expression can be induced by testosterone (final concentrations, 10 and 500 nM) and repressed by estrone sulphate (final concentration, 100 nM). Androstenone had no significant effect on SULT2B1 immunoreactive protein expression in the range of concentration, 10 nM to 1 μM. Time-courses (0 to 48 h) of steroid effects were investigated. The maximum effects of testosterone and estrone sulphate were observed in 24 h after the steroid treatments. This study provides direct evidence for involvement of sex steroids in the regulation of porcine hepatic SULTs.
- Published
- 2008
28. Human Hydroxysteroid Sulfotransferase SULT2B1 Pharmacogenomics: Gene Sequence Variation and Functional Genomics
- Author
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Irene Moon, Bruce W. Eckloff, Oreste E. Salavaggione, Jelena Zlatkovic, Eric D. Wieben, Daniel J. Schaid, Richard M. Weinshilboum, Bianca A. Thomae, and Yuan Ji
- Subjects
Gene isoform ,5' Flanking Region ,Leupeptins ,Recombinant Fusion Proteins ,Green Fluorescent Proteins ,Gene Expression ,Cysteine Proteinase Inhibitors ,Biology ,Linkage Disequilibrium ,White People ,chemistry.chemical_compound ,Exon ,Cytosol ,Gene Frequency ,Cell Line, Tumor ,SULT2B1 ,Chlorocebus aethiops ,Animals ,Humans ,Gene ,Pharmacology ,Genetics ,Microscopy, Confocal ,Polymorphism, Genetic ,Alternative splicing ,Sequence Analysis, DNA ,Molecular biology ,Black or African American ,Isoenzymes ,Open reading frame ,Haplotypes ,chemistry ,Pharmacogenetics ,COS Cells ,Mutation ,Molecular Medicine ,Hydroxysteroid ,Sulfotransferases ,Functional genomics ,Gene Deletion - Abstract
The human hydroxysteroid sulfotransferase (SULT) 2B1 gene is a member of the cytosolic SULT gene superfamily. The two SULT2B1 isoforms, SULT2B1a and SULT2B1b, are encoded by a single gene as a result of alternative transcription initiation and alternative splicing. SULT2B1b catalyzes the sulfonation of 3beta-hydroxysteroid hormones and cholesterol, whereas SULT2B1a preferentially catalyzes pregnenolone sulfonation. We used a genotype-to-phenotype approach to identify and characterize common sequence variation in SULT2B1. Specifically, we resequenced all exons, splice junctions, and approximately 2.5 kb of the 5'-flanking regions (FRs) for each isoform using 60 DNA samples each from African-American and Caucasian-American subjects. We observed 100 polymorphisms, including four nonsynonymous coding single nucleotide polymorphisms and one 6-base pair deletion-all within the "shared" region of the open reading frame. Functional genomic studies of the wild type (WT) and five variant allozymes for each isoform performed with a mammalian expression system showed that variant allozyme activities ranged from 64 to 88% of WT for SULT2B1a and from 76 to 98% for SULT2B1b. Relative levels of immunoreactive protein were similar to those for enzyme activity. Luciferase reporter gene constructs for 2.5 kb of the SULT2B1b 5'-FR displayed a cell line-dependent pattern of variation in activity. Finally, deletion of the proline-rich SULT2B1 carboxyl terminus resulted in intracellular protein aggregate formation and accelerated degradation of the truncated protein. These studies resulted in the identification of common SULT2B1 gene sequence variation, as well as insight into the effects of that variation on the function of this important steroid-metabolizing enzyme.
- Published
- 2007
29. Cloning, characterization and tissue expression of rat SULT2B1a and SULT2B1b steroid/sterol sulfotransferase isoforms: Divergence of the rat SULT2B1 gene structure from orthologous human and mouse genes
- Author
-
Osamu Hanyu, Atsushi Kohjitani, Charles A. Strott, and Hirotoshi Fuda
- Subjects
Gene isoform ,Sulfotransferase ,DNA, Complementary ,Recombinant Fusion Proteins ,Amino Acid Motifs ,Genetic Vectors ,Molecular Sequence Data ,Gene Expression ,Biology ,Substrate Specificity ,Rats, Sprague-Dawley ,Mice ,Exon ,Exon trapping ,SULT2B1 ,Genetics ,medicine ,Animals ,Humans ,Amino Acid Sequence ,RNA, Messenger ,Cloning, Molecular ,3' Untranslated Regions ,Gene ,Conserved Sequence ,Glutathione Transferase ,Sequence Homology, Amino Acid ,Reverse Transcriptase Polymerase Chain Reaction ,Exons ,General Medicine ,Molecular biology ,Rats ,Isoenzymes ,Kinetics ,RNA splicing ,Pregnenolone ,Sulfotransferases ,Nucleic Acid Amplification Techniques ,medicine.drug - Abstract
cDNAs for rat SULT2B1 steroid/sterol sulfotransferase isoforms were cloned, and the encoded proteins overexpressed, purified and characterized. The rat SULT2B1a isoform avidly sulfonates pregnenolone but poorly utilizes cholesterol as a substrate, whereas cholesterol is more efficiently sulfonated than pregnenolone by the SULT2B1b isoform; on the other hand, neither isoform sulfonates dehydroepiandrosterone to any significant degree. Real-time PCR revealed that SULT2B1a was only expressed in brain and testis, whereas SULT2B1b was mainly expressed in skin, intestine and kidney. The SULT2B1 gene is unique among steroid/sterol sulfotransferase genes in that it encodes for two isoforms as a result of an alternative exon I. Interestingly, whereas the orthologous human and mouse SULT2B1 gene structures are identical, the rat SULT2B1 gene structure diverges. Similar to human and mouse SULT2B1 genes the rat SULT2B1 gene consists of an alternative exon I; however, as a result of exonic rearrangement, the genic locations of exons IA and IB are reversed in the rat gene. Where exon IA is located downstream of exon IB in the human and mouse SULT2B1 genes, in the rat SULT2B1 gene exon IA is located upstream of exon IB. Furthermore, unlike the case with human and mouse SULT2B1 genes where differential splicing is necessitated since a portion of exon IA is fused with exon IB to complete the SULT2B1b mRNA, this step is not required with the rat gene.
- Published
- 2006
30. Crystal Structure of Human Cholesterol Sulfotransferase (SULT2B1b) in the Presence of Pregnenolone and 3′-Phosphoadenosine 5′-Phosphate
- Author
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Masahiko Negishi, Hirotoshi Fuda, Karen A. Lee, Lars C. Pedersen, Young C. Lee, and Charles A. Strott
- Subjects
Gene isoform ,Sulfotransferase ,Stereochemistry ,Dehydroepiandrosterone ,Substrate (chemistry) ,Cell Biology ,Biochemistry ,chemistry.chemical_compound ,chemistry ,SULT2B1 ,Pregnenolone ,medicine ,Transferase ,Hydroxysteroid ,Molecular Biology ,medicine.drug - Abstract
The gene for human hydroxysteroid sulfotransferase (SULT2B1) encodes two peptides, SULT2B1a and SULT2B1b, that differ only at their amino termini. SULT2B1b has a predilection for cholesterol but is also capable of sulfonating pregnenolone, whereas SULT2B1a preferentially sulfonates pregnenolone and only minimally sulfonates cholesterol. We have determined the crystal structure of SULT2B1a and SULT2B1b bound to the substrate donor product 3′-phosphoadenosine 5′-phosphate at 2.9 and 2.4 A, respectively, as well as SULT2B1b in the presence of the acceptor substrate pregnenolone at 2.3 A. These structures reveal a different catalytic binding orientation for the substrate from a previously determined structure of hydroxysteroid sulfotransferase (SULT2A1) binding dehydroepiandrosterone. In addition, the amino-terminal helix comprising residues Asp19 to Lys26, which determines the specificity difference between the SULT2B1 isoforms, becomes ordered upon pregnenolone binding, covering the substrate binding pocket.
- Published
- 2003
31. Sulphonation of dehydroepiandrosterone and neurosteroids: molecular cloning, expression, and functional characterization of a novel zebrafish SULT2 cytosolic sulphotransferase
- Author
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T. Govind Pai, Chau-Ching Liu, Takuya Sugahara, Ming-Cheh Liu, and Yuh-Shyong Yang
- Subjects
DNA, Complementary ,Cations, Divalent ,Sequence analysis ,Molecular Sequence Data ,Biology ,Molecular cloning ,Biochemistry ,Gene Expression Regulation, Enzymologic ,Substrate Specificity ,Cytosol ,SULT2B1 ,Complementary DNA ,Enzyme Stability ,Escherichia coli ,Amino Acid Sequence ,Cloning, Molecular ,Molecular Biology ,Peptide sequence ,Zebrafish ,Phylogeny ,chemistry.chemical_classification ,Cloning ,Base Sequence ,Temperature ,Dehydroepiandrosterone ,Sequence Analysis, DNA ,Cell Biology ,Hydrogen-Ion Concentration ,Zebrafish Proteins ,biology.organism_classification ,Molecular biology ,Recombinant Proteins ,Amino acid ,Kinetics ,chemistry ,Steroids ,Sulfonic Acids ,Sulfotransferases ,Research Article - Abstract
By searching the zebrafish EST (expressed-sequence tag) database, we have identified two partial cDNA clones encoding the 5' and 3' regions of a putative zebrafish sulphotransferase (ST). Using the reverse transcription-PCR technique, a full-length cDNA encoding this zebrafish ST was successfully cloned. Sequence analysis revealed that this novel zebrafish ST displays 44%, 43% and 40% amino acid identity with mouse SULT2B1, human SULT2B1b and human SULT2A1 ST respectively. This zebrafish ST therefore appears to belong to the SULT2 cytosolic ST gene family. Recombinant zebrafish ST, expressed using the pGEX-2TK prokaryotic expression system and purified from transformed Escherichia coli cells, migrated as a 34 kDa protein upon SDS/PAGE. Purified zebrafish ST displayed a strong sulphonating activity toward DHEA (dehydroepiandrosterone), with a optimum pH of 9.5. The enzyme also exhibited activities toward several neurosteroids with differential K(m) and V(max) values. A thermostability experiment revealed the enzyme to be relatively stable over a temperature range between 20 degrees C and 43 degrees C. Among ten different divalent metal cations tested, Fe2+ and Cd(2+ exhibited small, but significant, stimulatory effects, whereas Hg2+ and Cu2+ displayed considerably stronger inhibitory effects on the DHEA-sulphonating activity of the enzyme. These results constitute the first study on the molecular cloning, expression, and characterization of a zebrafish cytosolic SULT2 ST.
- Published
- 2003
32. Mutational Analysis of Human Hydroxysteroid Sulfotransferase SULT2B1 Isoforms Reveals That Exon 1B of the SULT2B1 Gene Produces Cholesterol Sulfotransferase, whereas Exon 1A Yields Pregnenolone Sulfotransferase
- Author
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Charles A. Strott, Young C. Lee, Hirotoshi Fuda, Chikara Shimizu, and Norman B. Javitt
- Subjects
Gene isoform ,Sulfotransferase ,DNA Mutational Analysis ,Genetic Vectors ,Molecular Sequence Data ,Oligonucleotides ,Biology ,Biochemistry ,Catalysis ,Exon ,chemistry.chemical_compound ,SULT2B1 ,medicine ,Humans ,Protein Isoforms ,Amino Acid Sequence ,Molecular Biology ,chemistry.chemical_classification ,Dose-Response Relationship, Drug ,Sequence Homology, Amino Acid ,Cholesterol sulfotransferase activity ,Exons ,Cell Biology ,Molecular biology ,Protein Structure, Tertiary ,Amino acid ,Kinetics ,Cholesterol ,chemistry ,Mutagenesis, Site-Directed ,Pregnenolone ,Hydroxysteroid ,Sulfotransferases ,Gene Deletion ,medicine.drug - Abstract
As a result of an alternative exon 1, the gene for human hydroxysteroid sulfotransferase (SULTB1) encodes for two peptides differing only at their amino termini. The SULT2B1b isoform preferentially sulfonates cholesterol. Conversely, the SULT2B1a isoform avidly sulfonates pregnenolone but not cholesterol. The outstanding structural feature that distinguishes the SULT2B1 isoforms from the prototypical SULT2A1 isozyme is the presence of extended amino- and carboxyl-terminal ends in the former. Investigating the functional significance of this unique characteristic reveals that removal of 53 amino acids from the relatively long carboxyl-terminal end that is common to both SULT2B1 isoforms has no effect on the catalytic activity of either isoform. On the other hand, removal of 23 amino acids from the amino-terminal end that is unique to SULT2B1b results in loss of cholesterol sulfotransferase activity, whereas removal of 8 amino acids from the amino-terminal end that is unique to SULT2B1a has no effect on pregnenolone sulfotransferase activity. Deletion analysis along with site-directed mutagenesis of SULT2B1b reveal that the amino acid segment 19-23 residues from the amino terminus and particularly isoleucines at positions 21 and 23 are crucial for cholesterol catalysis. In the gene for SULT2B1, exon 1B encodes for only the unique amino-terminal region of SULT2B1b; however, exon 1A encodes for the unique amino-terminal end of SULT2B1a plus an additional 48 amino acids. Thus, if the gene for SULT2B1 employs exon 1B, cholesterol sulfotransferase is synthesized, whereas if exon 1A is used, pregnenolone sulfotransferase is produced.
- Published
- 2002
33. Identification of key genes affecting disease free survival time of pediatric acute lymphoblastic leukemia based on bioinformatic analysis
- Author
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Jing-Hua Wang, Xi Chen, Xin-Guo Luo, and Haiyan Gao
- Subjects
Male ,Candidate gene ,Disease free survival ,Key genes ,Adolescent ,Biology ,Disease-Free Survival ,Pediatric Acute Lymphoblastic Leukemia ,SULT2B1 ,Humans ,Child ,Molecular Biology ,Gene ,Genetics ,Gene Expression Regulation, Leukemic ,Computational Biology ,Infant ,Cell Biology ,Hematology ,Precursor Cell Lymphoblastic Leukemia-Lymphoma ,Neoplasm Proteins ,Survival Rate ,Differentially expressed genes ,Child, Preschool ,Molecular Medicine ,Identification (biology) ,Female ,Genes, Neoplasm - Abstract
The poor prognosis of pediatric acute lymphoblastic leukemia (ALL) indicates the existence of key candidate genes that affect pediatric ALL and its prognosis. The limma package in R was applied to screen differentially expressed genes (DEGs), and the Survival package and KMsurv package in R were used to screen disease free survival time related genes (prognosis genes). Then, based on latent pathway identification analysis (LPIA), latent pathways were identified, and pathway–pathway interaction network was constructed and visualized by Cytoscape. Based on the expression values of 8284 genes in 126 chips, 2796 DEGs and 353 prognosis genes were screened out. After overlapping DEGs and prognosis genes, 75 key genes were identified, which were most significantly enriched in 25 GO functions and chronic myeloid leukemia pathway. For the 75 key genes, 27 disease risk sub-pathways were identified, and HK3 , HNMT , SULT2B1 , KYNU , and PTGS2 were the significant key genes which were enriched in these sub-pathways. Furthermore, based on pathway–pathway interaction analysis, HK3 and PTGS2 were predicted as the most important genes. Through glycolysis and arachidonic acid metabolism, HK3 and PTGS2 might play important roles in pediatric ALL and its prognosis, and thus, might be potential targets for therapeutic intervention to suppress pediatric ALL.
- Published
- 2014
34. Biochemical Characterization and Tissue Distribution of Human SULT2B1
- Author
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William J. Geese and Rebecca Blanchard Raftogianis
- Subjects
Sulfotransferase ,Molecular Sequence Data ,Biophysics ,Dehydroepiandrosterone ,Biology ,Biochemistry ,Substrate Specificity ,chemistry.chemical_compound ,Sulfation ,SULT2B1 ,Enzyme Stability ,medicine ,Humans ,Tissue Distribution ,Amino Acid Sequence ,RNA, Messenger ,Molecular Biology ,Hydroxysteroids ,chemistry.chemical_classification ,Sequence Homology, Amino Acid ,Dihydrotestosterone ,Stereoisomerism ,Cell Biology ,Recombinant Proteins ,In vitro ,Kinetics ,Enzyme ,chemistry ,Hydroxysteroid ,Sulfotransferases ,medicine.drug - Abstract
The human hydroxysteroid sulfotransferase (SULT) family is comprised of two subfamilies, SULT2A1 and SULT2B1. We characterized the substrate specificity, in vitro biochemical properties, and tissue distribution patterns of human SULT2B1a and SULT2B1b. In contrast to the wide substrate specificity of SULT2A1, SULT2B1a and SULT2B1b specifically catalyzed the sulfonation of 3β-hydroxysteroids with high catalytic efficiency. Both SULT2B1 enzymes also sulfonated dihydrotestosterone. In vitro studies revealed that the biochemical properties of SULT2B1a and SULT2B1b were not significantly different from each other. However, tissue expression analysis suggested that they are differentially regulated. In contrast to the limited tissue distribution of SULT2A1, SULT2B1 was detected in a variety of hormone-responsive tissues including placenta, ovary, uterus, and prostate. The catalytic activity toward dehydroepiandrosterone and dihydrotestosterone, biologically important androgens, coupled with expression in prostate suggests that SULT2B1 may play a novel regulatory role that protects against the mitogenic effects of androgens.
- Published
- 2001
35. Human Hydroxysteroid Sulfotransferase SULT2B1: Two Enzymes Encoded by a Single Chromosome 19 Gene
- Author
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Louis S. Ramagli, Evan E. Eichler, Michael J. Siciliano, Harvey W. Mohrenweiser, Thomas C. Wood, Richard M. Weinshilboum, and Chengtao Her
- Subjects
Male ,DNA, Complementary ,Placenta ,Molecular Sequence Data ,Biology ,Substrate Specificity ,Exon ,chemistry.chemical_compound ,Pregnancy ,SULT2B1 ,Genetics ,Animals ,Humans ,Amino Acid Sequence ,RNA, Messenger ,Cloning, Molecular ,Gene ,Peptide sequence ,DNA Primers ,chemistry.chemical_classification ,Base Sequence ,Alternative splicing ,Prostate ,Nucleic acid sequence ,Chromosome Mapping ,Molecular biology ,Recombinant Proteins ,Amino acid ,Trachea ,Alternative Splicing ,Biochemistry ,chemistry ,COS Cells ,Female ,Hydroxysteroid ,Sulfotransferases ,Chromosomes, Human, Pair 19 - Abstract
We have cloned and characterized cDNAs that encode two human hydroxysteroid sulfotransferase (SULT) enzymes, SULT2B1a and SULT2B1b, as well as the single gene that encodes both of these enzymes. The two cDNAs differed at their 5'-termini and had 1050- and 1095-bp open reading frames that encoded 350 and 365 amino acids, respectively. The amino acid sequences encoded by these cDNAs included "signature sequences" that are conserved in all known cytosolic SULTs. Both cDNAs appeared, on the basis of amino acid sequence analysis, to be members of the hydroxysteroid SULT "family, " SULT2, but they were only 48% identical in amino acid sequence with the single known member of that family in humans, SULT2A1 (also referred to as DHEA ST). Northern blot analysis demonstrated the presence of SULT2B1 mRNA species approximately 1.4 kb in length in human placenta, prostate, and trachea and-faintly-in small intestine and lung. Expression of the two human SULT2B1 cDNAs in COS-1 cells showed that both of the encoded proteins catalyzed sulfation of the prototypic hydroxysteroid SULT substrate, dehydroepiandrosterone, but both failed to catalyze the sulfate conjugation of 4-nitrophenol or 17beta-estradiol, prototypic substrates for the phenol and estrogen SULT subfamilies. Both of these cDNAs were encoded by a single gene, SULT2B1. The locations of most exon-intron splice junctions in SULT2B1 were identical to those of the only other known human hydroxysteroid SULT gene SULT2A1 (previously STD). The divergence in 5'-terminal sequences of the two SULT2B1 cDNAs resulted from alternative transcription initiation prior to different 5' exons, combined with alternative splicing. SULT2B1 mapped to human chromosome band 19q13.3, approximately 500 kb telomeric to the location of SULT2A1.
- Published
- 1998
36. SULT2B1: unique properties and characteristics of a hydroxysteroid sulfotransferase family
- Author
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Katie Jo Rohn-Glowacki and Charles N. Falany
- Subjects
Gene isoform ,Proline ,Molecular Sequence Data ,Sequence alignment ,Biology ,Isozyme ,Sulfation ,Cytosol ,Species Specificity ,SULT2B1 ,Serine ,Gene family ,Animals ,Humans ,Pharmacology (medical) ,Amino Acid Sequence ,General Pharmacology, Toxicology and Pharmaceutics ,Gene ,Peptide sequence ,Cell Nucleus ,Isoenzymes ,Molecular Docking Simulation ,Cholesterol ,Biochemistry ,Sulfotransferases ,Sequence Alignment ,Protein Binding - Abstract
The SULT2b gene family consists of a single gene capable of generating two functional transcripts utilizing different transcriptional start sites in the first exon. This results in the translation of two separate proteins, SULT2B1a and SULT2B1b, with different amino-terminal peptides and approximately 95% identical sequences. The second distinguishing feature of the SULT2B isoforms is the proline/serine-rich carboxy-terminal sequence. To date, presence of the SULT2B gene appears limited to mammals and there is also only limited conservation of structure or sequence of the carboxy-terminal peptide. Although both SULT2B1 messages are present in human tissues, to date, only the SULT2B1b protein has been detected in the tissues investigated. In contrast, selective expression of SULT2B1a has been detected in rodent brain, whereas SULT2B1b was expressed in skin and intestine. Characterization of the SULT2B1 isoforms has been limited by the inability to isolate reliably active SULT2B1b from tissues or cells. SULT2B1 cDNAs can be expressed in Escherichia coli and the expressed active enzymes show selectivity for sulfation of 3β-hydroxysteroids. SULT2B1b due to the binding properties of the amino-terminal peptides also shows high cholesterol sulfation activity. Although human SULT2B1b displays significant substrate cross-reactivity with SULT2A1, the isoforms have different tissue expression patterns. Human SULT2B1b also shows nuclear localization in selected tissues that appears related to serine phosphorylation of the carboxy-terminal peptide. Overall, the understanding of the properties and function of the SULT2B1 isoforms is limited and the structural variability of the unique amino- and carboxy-sequences suggests significant species differences that need to be investigated.
- Published
- 2013
37. 5,6-Epoxy-cholesterols contribute to the anticancer pharmacology of Tamoxifen in breast cancer cells
- Author
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Gregory, Segala, Medina, P. D., De Medina, P., Philippe Medina, D. E., Iuliano, Luigi, Zerbinati, Chiara, Paillasse, Michael R., Emmanuel, Noguer, Florence, Dalenc, Bruno, Payre, Craig Jordan, V., Michel, Record, Sandrine Silvente Poirot, Marc, Poirot, Centre de Recherches en Cancérologie de Toulouse (CRCT), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Claudius Regaud, CRLCC Institut Claudius Regaud, Université Fédérale Toulouse Midi-Pyrénées, Affichem, Department of Medico-Surgical Sciences & Biotechnologies, Università degli Studi di Roma 'La Sapienza' [Rome], Centre de Microscopie Électronique Appliquée à la Biologie (CMEAB), Hôpital de Rangueil, CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse]-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Lombardi comprehensive cancer center, Georgetown University Medical Center, Sterol metabolism and therapeutic innovations in oncology, CRLCC Institut Claudius Regaud-CRLCC Institut Claudius Regaud-Université Toulouse III - Paul Sabatier (UT3), Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse]-Toulouse Réseau Imagerie-Genotoul ( TRI-Genotoul), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut Claudius Regaud, Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Poirot, Marc, Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome] (UNIROMA), and Toulouse Réseau Imagerie-Genotoul ( TRI-Genotoul)
- Subjects
Selective Estrogen Receptor Modulators ,aebs ,Pyrrolidines ,Oxysterol ,breast cancer ,cheh ,sulfation ,cholesterol epoxide ,Antineoplastic Agents ,Breast Neoplasms ,Pharmacology ,Ligands ,Biochemistry ,03 medical and health sciences ,0302 clinical medicine ,Estrogen Receptor Modulators ,SULT2B1 ,Cell Line, Tumor ,medicine ,Humans ,Liver X receptor ,Receptor ,Triglycerides ,030304 developmental biology ,Liver X Receptors ,Epoxide Hydrolases ,0303 health sciences ,Binding Sites ,Chemistry ,[SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences ,Antiestrogen ,Orphan Nuclear Receptors ,3. Good health ,[SDV.SP] Life Sciences [q-bio]/Pharmaceutical sciences ,Tamoxifen ,Cholesterol ,Selective estrogen receptor modulator ,030220 oncology & carcinogenesis ,Female ,Signal transduction ,Sulfotransferases ,Reactive Oxygen Species ,Oxidation-Reduction ,medicine.drug - Abstract
International audience; Tamoxifen (Tam) is a selective estrogen receptor modulator (SERM) that remains one of the major drugs used in the hormonotherapy of breast cancer (BC). In addition to its SERM activity, we recently showed that the oxidative metabolism of cholesterol plays a role in its anticancer pharmacology. We established that these effects were not regulated by the ER but by the microsomal antiestrogen binding site/cholesterol-5,6-epoxide hydrolase complex (AEBS/ChEH). The present study aimed to identify the oxysterols that are produced under Tam treatment and to define their mechanisms of action. Tam and PBPE (a selective AEBS/ChEH ligand) stimulated the production and the accumulation of 5,6α-epoxy-cholesterol (5,6α-EC), 5,6α-epoxy-cholesterol-3β-sulfate (5,6-ECS), 5,6β-epoxy-cholesterol (5,6β-EC) in MCF-7 cells through a ROS-dependent mechanism, by inhibiting ChEH and inducing sulfation of 5,6α-EC by SULT2B1b. We showed that only 5,6α-EC was responsible for the induction of triacylglycerol (TAG) biosynthesis by Tam and PBPE, through the modulation of the oxysterol receptor LXRβ. The cytotoxicity mediated by Tam and PBPE was triggered by 5,6β-EC through an LXRβ-independent route and by 5,6-ECS through an LXRβ-dependent mechanism. The importance of SULT2B1b was confirmed by its ectopic expression in the SULT2B1b(-) MDA-MB-231 cells, which became sensitive to 5,6α-EC, Tam or PBPE at a comparable level to MCF-7 cells. This study established that 5,6-EC metabolites contribute to the anticancer pharmacology of Tam and highlights a novel signaling pathway that points to a rationale for re-sensitizing BC cells to Tam and AEBS/ChEH ligands.
- Published
- 2013
38. Hydroxysteroid sulfotransferase SULT2B1b promotes hepatocellular carcinoma cells proliferation in vitro and in vivo
- Author
-
Lianhua Yin, Yanxia Ning, Yali Xu, Xiaoming Yang, Xiaobo Li, Fenghua Guo, and Xiuling Zhi
- Subjects
Cell cycle checkpoint ,Anatomy and Physiology ,Digestive Physiology ,lcsh:Medicine ,Gene Expression ,Apoptosis ,Mice ,Nude mouse ,Molecular Cell Biology ,Gastrointestinal Cancers ,RNA, Small Interfering ,lcsh:Science ,Multidisciplinary ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Liver Neoplasms ,Flow Cytometry ,Gene Expression Regulation, Neoplastic ,Medicine ,RNA Interference ,Sulfotransferases ,Research Article ,Cell Physiology ,Carcinoma, Hepatocellular ,Blotting, Western ,Gastroenterology and Hepatology ,Real-Time Polymerase Chain Reaction ,Cell Growth ,In vivo ,SULT2B1 ,Cell Line, Tumor ,Genetics ,Animals ,Humans ,Biology ,Cell Proliferation ,DNA Primers ,Analysis of Variance ,Cell growth ,lcsh:R ,Cell Cycle Checkpoints ,biology.organism_classification ,Molecular biology ,In vitro ,digestive system diseases ,Microscopy, Fluorescence ,Cell culture ,Genetics of Disease ,lcsh:Q ,Gene Function ,Digestive System - Abstract
Hydroxysteroid sulfotransferase 2B1b (SULT2B1b) is highly selective for the addition of sulfate groups to 3β-hydroxysteroids. Although previous reports have suggested that SULT2B1b is correlated with cell proliferation of hepatocytes, the relationship between SULT2B1b and the malignant phenotype of hepatocarcinoma cells was not clear. In the present study, we found that SULT2B1 was comparatively higher in the human hepatocarcinoma tumorous tissues than their adjacent tissues. Besides, SULT2B1b overexpression promoted the growth of the mouse hepatocarcinoma cell line Hepa1-6, while Lentivirus-mediated SULT2B1b interference inhibited growth as assessed by the CCK-8 assay. Likewise, inhibition of SULT2B1b expression induced cell-cycle arrest and apoptosis in Hepa1-6 cells by upregulating the expression of FAS, downregulating the expression of cyclinB1, BCL2 and MYC in vitro and in vivo at both the transcript and protein levels. Knock-down of SULT2B1b expression significantly suppressed tumor growth in nude mouse xenografts. Moreover, proliferation rates and SULT2B1b expression were highly correlated in the human hepatocarcinoma cell lines Huh-7, Hep3B, SMMC-7721 and BEL-7402 cells. Knock-down of SULT2B1b inhibited cell growth and cyclinB1 levels in human hepatocarcinoma cells and suppressed xenograft growth in vivo. In conclusion, SULT2B1b expression promotes proliferation of hepatocellular carcinoma cells in vitro and in vivo, which may contribute to the progression of HCC.
- Published
- 2012
39. Disturbed balance between phase I and II metabolizing enzymes in ovarian endometriosis: a source of excessive hydroxy-estrogens and ROS?
- Author
-
Neli Hevir, Tea Lanišnik Rižner, and Martina Ribič-Pucelj
- Subjects
Adult ,medicine.medical_specialty ,medicine.drug_class ,Blotting, Western ,Endometriosis ,Ovary ,Biology ,Catechol O-Methyltransferase ,Biochemistry ,Isozyme ,Gene Expression Regulation, Enzymologic ,Young Adult ,Endocrinology ,Downregulation and upregulation ,Internal medicine ,SULT2B1 ,medicine ,Cytochrome P-450 CYP1A1 ,NAD(P)H Dehydrogenase (Quinone) ,Humans ,Molecular Biology ,Catechol-O-methyl transferase ,Gene Expression Profiling ,Estrogens ,medicine.disease ,Immunohistochemistry ,Metabolic Detoxication, Phase II ,Up-Regulation ,Isoenzymes ,medicine.anatomical_structure ,Glutathione S-Transferase pi ,Steroid 16-alpha-Hydroxylase ,Estrogen ,Case-Control Studies ,Cytochrome P-450 CYP1B1 ,Steroid Hydroxylases ,Ovarian Endometriosis ,Female ,Metabolic Detoxication, Phase I ,Aryl Hydrocarbon Hydroxylases ,Reactive Oxygen Species - Abstract
Oxidative metabolism of estrogens was studied in 31 ovarian endometriosis and 29 normal endometrium samples, by qPCR. Expression was monitored for genes encoding five estrogen hydroxylating, five hydroxy (OH)-estrogen conjugating, and three estrogen quinone detoxifying enzymes. CYP1B1, COMT, NQO1, and GSTP1 protein levels were determined using Western blotting and immunohistochemistry staining. Increased expression of CYP1A1, CYP3A7 and COMT, and higher levels of MB-COMT were seen in endometriosis, as compared to normal endometrium. Expression of CYP1B1, CYP3A5, SULT1A1 and NQO2 was unchanged, with comparable CYP1B1 protein levels. Expression of SULT1E1, SULT2B1, UGT2B7, NQO1, and GSTP1 was decreased. Three NQO1 isoforms were detected; NQO1c appears to be endometriosis-specific. Our data indicate a disturbed balance between phase I and II metabolizing enzymes in endometriosis, potentially leading to excessive OH-estrogen and altered ROS formation, and stimulation of proliferation of ectopic endometrium. This is the first report on disturbed expression of estrogen oxidative metabolism genes in ovarian endometriosis.
- Published
- 2012
40. Cholesterol-5,6-epoxides: chemistry, biochemistry, metabolic fate and cancer
- Author
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Sandrine Silvente-Poirot, Marc Poirot, Métabolisme du cholestérol et innovations thérapeutiques en oncologie, Institut Claudius Regaud, and Poirot, Marc
- Subjects
Cholesterol ,Cancer ,General Medicine ,Metabolism ,[SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences ,medicine.disease ,Biochemistry ,[SDV.SP] Life Sciences [q-bio]/Pharmaceutical sciences ,chemistry.chemical_compound ,chemistry ,SULT2B1 ,Detoxification ,Neoplasms ,Terminology as Topic ,Epoxide Hydrolases ,medicine ,Animals ,Humans ,Tamoxifen ,Carcinogen ,medicine.drug - Abstract
International audience; In the nineteen sixties it was proposed that cholesterol might be involved in the etiology of cancers and cholesterol oxidation products were suspected of being causative agents. Researchers had focused their attention on cholesterol-5,6-epoxides (5,6-ECs) based on several lines of evidence: 1) 5,6-ECs contained an oxirane group that was supposed to confer alkylating properties such as those observed for aliphatic and aromatic epoxides. 2) cholesterol-5,6-epoxide hydrolase (ChEH) was induced in pre-neoplastic lesions of skin from rats exposed to ultraviolet irradiations and ChEH was proposed to be involved in detoxification processes like other epoxide hydrolases. However, 5,6-ECs failed to induce carcinogenicity in rodents which ruled out a potent carcinogenic potential for 5,6-ECs. Meanwhile, clinical studies revealed an anomalous increase in the concentrations of 5,6β-EC in the nipple fluids of patients with pre-neoplastic breast lesions and in the blood of patients with endometrious cancers, suggesting that 5,6-ECs metabolism could be linked with cancer. Paradoxically, ChEH has been recently shown to be totally inhibited by therapeutic concentrations of tamoxifen (Tam), which is one of the main drugs used in the hormonotherapy and the chemoprevention of breast cancers. These data would suggest that the accumulation of 5,6-ECs could represent a risk factor, but we found that 5,6-ECs were involved in the induction of breast cancer cell differentiation and death induced by Tam suggesting a positive role of 5,6-ECs. These observations meant that the biochemistry and the metabolism of 5,6-ECs needed to be extensively studied. We will review the current knowledge and the future direction of 5,6-ECs chemistry, biochemistry, metabolism, and relationship with cancer.
- Published
- 2012
41. Human cytosolic sulfotransferase 2B1: isoform expression, tissue specificity and subcellular localization
- Author
-
Charles N. Falany, Nicole A. Dumas, Josie L. Falany, Andra R. Frost, and Dongning He
- Subjects
Gene isoform ,Sulfotransferase ,Transcription, Genetic ,Endocrinology, Diabetes and Metabolism ,Placenta ,Clinical Biochemistry ,Molecular Sequence Data ,Breast Neoplasms ,Biology ,Biochemistry ,Article ,Substrate Specificity ,Mice ,Endocrinology ,Sulfation ,Cytosol ,SULT2B1 ,medicine ,Animals ,Humans ,Protein Isoforms ,Amino Acid Sequence ,Breast ,Choriocarcinoma ,Cloning, Molecular ,Molecular Biology ,Skin ,Cell Nucleus ,Sequence Homology, Amino Acid ,Sulfates ,Cell Biology ,Dehydroepiandrosterone ,Subcellular localization ,Rats ,Cell nucleus ,medicine.anatomical_structure ,Carcinoma, Intraductal, Noninfiltrating ,Cholesterol ,Gene Expression Regulation ,Pregnenolone ,Molecular Medicine ,Sulfotransferases ,Nuclear localization sequence ,medicine.drug ,Subcellular Fractions - Abstract
Sulfation is an important Phase II conjugation reaction involved in the synthesis and metabolism of steroids in humans. Two different isoforms (2B1a and 2B1b) are encoded by the sulfotransferase (SULT) 2B1 gene utilizing different start sites of transcription resulting in the incorporation of different first exons. SULT2B1a and SULT2B1b are 350 and 365 amino acids in length, respectively, and the last 342 aa are identical. Message for both SULT2B1 isoforms is present in human tissues although SULT2B1b message is generally more abundant. However, to date only SULT2B1b protein has been detected in human tissues or cell lines. SULT2B1b is localized in the cytosol and/or nuclei of human cells. A unique 3'-extension of SULT2B1b is required for nuclear localization in human BeWo placental choriocarcinoma cells. Nuclear localization is stimulated by forskolin treatment in BeWo cells and serine phosphorylation has been identified in the 3'-extension. SULT2B1b is selective for the sulfation of 3beta-hydroxysteroids such as dehydroepiandrosterone and pregnenolone, and may also have a role in cholesterol sulfation in human skin. The substrate specificity, nuclear localization, and tissue localization of SULT2B1b suggest a role in regulating the responsiveness of cells to adrenal androgens via their direct inactivation or by preventing their conversion to more potent androgens and estrogens.
- Published
- 2006
42. Cholesterol sulfotransferase (Sult2b1) inactivates oxysterol ligands of LXR
- Author
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Wenling Chen, David J. Mangelsdorf, Guoxun Chen, and David W. Russell
- Subjects
Oxysterol ,Biochemistry ,Chemistry ,SULT2B1 ,Genetics ,Cholesterol sulfotransferase ,Liver X receptor ,Molecular Biology ,Biotechnology - Published
- 2006
43. SULFATION PATHWAYS: Expression of SULT2A1 , SULT2B1 and HSD3B1 in the porcine testis and epididymis.
- Author
-
Zimmer B, Tenbusch L, Klymiuk MC, Dezhkam Y, and Schuler G
- Subjects
- 17-Hydroxysteroid Dehydrogenases metabolism, 3-Oxo-5-alpha-Steroid 4-Dehydrogenase metabolism, Animals, Aromatase metabolism, Cholesterol Side-Chain Cleavage Enzyme metabolism, Hydroxysteroids metabolism, Leydig Cells metabolism, Male, Steroid 17-alpha-Hydroxylase, Steroid Isomerases metabolism, Sulfotransferases metabolism, Swine, Epididymis metabolism, Sulfates metabolism, Testis metabolism
- Abstract
In the porcine testis, in addition to estrogen sulfates, the formation of numerous sulfonated neutral hydroxysteroids has been observed. However, their functions and the underlying synthetic pathways are still widely unclear. To obtain further information on their formation in postpubertal boars, the expression of sulfotransferases considered relevant for neutral hydroxysteroids ( SULT2A1, SULT2B1 ) was investigated in the testis and defined segments of the epididymis applying real-time RT-qPCR, Western blot and immunohistochemistry (IHC). Sulfotransferase activities were assessed in tissue homogenates or cytosolic preparations applying dehydroepiandrosterone and pregnenolone as substrates. A high SULT2A1 expression was confirmed in the testis and localized in Leydig cells by IHC. In the epididymis, SULT2A1 expression was virtually confined to the body. SULT2B1 expression was absent or low in the testis but increased significantly along the epididymis. Immunohistochemical observations indicate that both enzymes are secreted into the ductal lumen via an apocrine mechanism. The results from the characterization of expression patterns and activity measurements suggest that SULT2A1 is the prevailing enzyme for the sulfonation of hydroxysteroids in the testis, whereas SULT2B1 may catalyze the formation of sterol sulfates in the epididymis. In order to obtain information on the overall steroidogenic capacity of the porcine epididymis, the expression of important steroidogenic enzymes ( CYP11A1, CYP17A1, CYP19, HSD3B1, HSD17B3, SRD5A2 ) was monitored in the defined epididymal segments applying real-time RT-qPCR. Surprisingly, in addition to a high expression of SRD5A2 in the epididymal head, a substantial expression of HSD3B1 was detected, which increased along the organ., (© 2018 Society for Endocrinology.)
- Published
- 2018
- Full Text
- View/download PDF
44. Identification and immunohistochemical localization of Sulfotransferase 2B1b (SULT2B1b) in human lung
- Author
-
Dongning He, Andra R. Frost, and Charles N. Falany
- Subjects
Gene isoform ,Sulfotransferase ,Alternative splicing ,Biophysics ,Bronchi ,Epithelial Cells ,Biology ,Biochemistry ,Molecular biology ,Immunohistochemistry ,Alveolar cells ,Sulfation ,medicine.anatomical_structure ,SULT2B1 ,Pregnenolone ,medicine ,Humans ,Northern blot ,RNA, Messenger ,Sulfotransferases ,Molecular Biology ,Lung ,medicine.drug - Abstract
Sulfotransferase 2B1b (SULT2B1b) is a member of SULT 2 gene family. SULT2B1a and SULT2B1b are transcribed from the same gene using different transcriptional start sites and contain different first exons as the result of alternative splicing. SULT2B1a and SULT2B1b are 350 and 365 AA in length, respectively. Northern blot analysis and SULT2B1 isoform specific RT-PCR detected only SULT2B1b message in human lung tissue. Immunoblot analysis of human lung tissue with a specific rabbit anti-SULT2B1 polyclonal antibody detected only SULT2B1b immunoreactive protein in human lung cytosol. Immunoprecipitation and MALDI mass spectroscopic analysis verified that the immunoreactive protein was SULT2B1b. Immunohistochemical localization of SULT2B1b in human tissues showed expression in the cytoplasm of ciliated columnar or cuboidal epithelial cells in terminal bronchia. No staining was observed in alveolar cells. SULT2B1b is selective for the sulfation of 3beta-hydroxysteroids such as dehydroepiandrosterone and pregnenolone as well as cholesterol. The presence of SULT2B1b in lung tissues suggests a role in the regulation of local steroid hormone synthesis and metabolism.
- Published
- 2004
45. Steroid sulfotransferase 2A1 gene transcription is regulated by steroidogenic factor 1 and GATA-6 in the human adrenal
- Author
-
Karla J. Saner, Bruce R. Carr, Hironobu Sasano, Takashi Suzuki, William E. Rainey, Jerome F. Strauss, Clement K M Ho, and John Pizzey
- Subjects
Steroidogenic factor 1 ,Transcriptional Activation ,endocrine system ,Sulfotransferase ,Transcription, Genetic ,Dehydroepiandrosterone ,Receptors, Cytoplasmic and Nuclear ,Electrophoretic Mobility Shift Assay ,Biology ,Response Elements ,Steroidogenic Factor 1 ,Cell Line ,chemistry.chemical_compound ,Endocrinology ,Dehydroepiandrosterone sulfate ,Sulfation ,SULT2B1 ,GATA6 Transcription Factor ,Adrenal Glands ,Humans ,Protein Isoforms ,Steroid sulfotransferase ,RNA, Messenger ,Promoter Regions, Genetic ,Molecular Biology ,Transcription factor ,Homeodomain Proteins ,Binding Sites ,General Medicine ,DNA-Binding Proteins ,chemistry ,Biochemistry ,Gene Expression Regulation ,Organ Specificity ,Sulfotransferases ,Transcription Factors - Abstract
Sulfonation is a phase II conjugation reaction responsible for the biotransformation of many compounds including steroids, bile acids, and drugs. Humans are presently known to express at least five cytosolic sulfotransferase (SULT) enzymes, of which only two are hydroxysteroid SULT, SULT2A1, commonly known as steroid sulfotransferase, and the cholesterol sulfotransferase SULT2B1. SULT2A1 is highly expressed in the adrenal where it is responsible for the sulfation of hydroxysteroids including conversion of dehydroepiandrosterone to dehydroepiandrosterone sulfate and in the liver where it is responsible for sulfation of bile acids and circulating hydroxysteroids. Little is known concerning the transcriptional regulation of human SULT2A1 in adrenal. Herein we demonstrate the role of two transcription factors, steroidogenic factor 1 (SF1) and GATA-6, in the regulation of SULT2A1 transcription. These transcription factors were quantified by real-time RT-PCR in normal human adrenal tissue. Transient transfection assays with deleted and mutated SULT2A1 promoter constructs allowed for the determination of specific SF1 and GATA binding cis-regulatory elements necessary for transactivation of SULT2A1 promoter, and binding was confirmed by EMSA analysis. Both SF1 and GATA-6 were positive regulators of SULT2A1 promoter constructs. These data support the hypothesis that adrenal SULT2A1 expression is regulated by SF1 and GATA-6.
- Published
- 2004
46. Conservation of the hydroxysteroid sulfotransferase SULT2B1 gene structure in the mouse: pre- and postnatal expression, kinetic analysis of isoforms, and comparison with prototypical SULT2A1
- Author
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Charles A. Strott, Hidekatsu Yanai, Hirotoshi Fuda, and Chikara Shimizu
- Subjects
Gene isoform ,Sulfotransferase ,Reverse Transcriptase Polymerase Chain Reaction ,Molecular Sequence Data ,Gene Expression Regulation, Developmental ,Biology ,Molecular biology ,Gene Expression Regulation, Enzymologic ,Mice, Inbred C57BL ,Exon ,chemistry.chemical_compound ,Kinetics ,Mice ,Endocrinology ,chemistry ,SULT2B1 ,Complementary DNA ,RNA splicing ,Animals ,Female ,Hydroxysteroid ,Amino Acid Sequence ,Cloning, Molecular ,Sulfotransferases ,Gene - Abstract
A novel mouse hydroxysteroid sulfotransferase cDNA has been cloned, and organization of its gene structure has been determined. The new mouse sulfotransferase, SULT2B1a, and its closely related isoform, SULT2B1b, are derived from a single SULT2B1 gene as a result of an alternative exon I and differential splicing. Thus, the only structural distinction between the two SULT2B1 isoforms is at their amino-terminal ends. Importantly, in contrast to the prototypical mouse hydroxysteroid sulfotransferase SULT2A1, the SULT2B1 isoforms have a predilection for cholesterol. Real-time RT-PCR reveals that the SULT2B1a isoform is most abundantly expressed in the brain and spinal cord, whereas SULT2B1b and SULT2A1 are weakly, if at all, expressed in the central nervous system. On the other hand, the SULT2B1b isoform is the most prominent hydroxysteroid sulfotransferase expressed in skin, whereas SULT2A1 is strikingly expressed in the liver. The substrate specificities and differential expression patterns of the three SULT2 isozymes strongly suggest that they have distinct biologic roles to play. Of further interest, the mouse SULT2B1 and SULT2A1 genes are differentially expressed during embryonic development, with the former being expressed at all stages from E8.5-E19, whereas the latter is not expressed until E19. It is speculated that, during embryonic development, SULT2B1b is required for production of cholesterol sulfate essential for normal skin development, whereas SULT2B1a produces pregnenolone sulfate, an essential neurosteroid during development of the central nervous system.
- Published
- 2003
47. Expression and characterization of the human 3 beta-hydroxysteroid sulfotransferases (SULT2B1a and SULT2B1b)
- Author
-
Connie A. Meloche and Charles N. Falany
- Subjects
Sulfotransferase ,Endocrinology, Diabetes and Metabolism ,Clinical Biochemistry ,Immunoblotting ,Molecular Sequence Data ,Androstenediol ,Epiandrosterone ,Alcohol sulfotransferase ,Biochemistry ,chemistry.chemical_compound ,Endocrinology ,Sulfation ,SULT2B1 ,medicine ,Escherichia coli ,Humans ,Amino Acid Sequence ,Cloning, Molecular ,Molecular Biology ,biology ,Sequence Homology, Amino Acid ,Cell Biology ,Recombinant Proteins ,Kinetics ,chemistry ,biology.protein ,Pregnenolone ,Molecular Medicine ,Hydroxysteroid ,Sulfotransferases ,hormones, hormone substitutes, and hormone antagonists ,medicine.drug - Abstract
The human hydroxysteroid sulfotransferase, dehydroepiandrosterone sulfotransferase (DHEA-ST), is highly expressed in liver and adrenal cortex and displays reactivity towards a broad range of hydroxysteroids including 3 beta-hydroxysteroids, 3 alpha-hydroxysteroids, estrogens with a 3-phenolic moiety, and 17-hydroxyl group of androgens. In contrast, characterization of the newly described human hydroxysteroid sulfotransferase SULT2B1 isoforms shows that these enzymes are selective for the sulfation of 3 beta-hydroxysteroids, such as pregnenolone, epiandrosterone, DHEA, and androstenediol. There was no activity detected towards testosterone, dexamethasone, beta-estradiol, androsterone, or p-nitrophenol. The SULT2B1 gene encodes two isoforms, SULT2B1a and SULT2B1b, which are generated by alternate splicing of the first exon; therefore the SULT2B1 isoforms differ at their N-terminals. Northern Blot analysis detected a SULT2B1 message in RNA isolated from the human prostate and placenta. No SULT2B1 message was observed in RNA isolated from human liver, colon, lung, kidney, brain, or testis tissue. Purified SULT2B1a was used to generate a specific rabbit polyclonal anti-SULT2B1 antibody. The anti-SULT2B1 antibody did not react with expressed human EST, P-PST-1, M-PST, DHEA-ST, or ST1B2, during immunoblot analysis. The substrate specificity of the expressed SULT2B1 isoforms suggests that these enzymes are capable of regulating the activity of adrenal androgens in human tissues via their inactivation by sulfation.
- Published
- 2001
48. Molecular cloning, expression, and functional characterization of novel mouse sulfotransferases
- Author
-
Yoichi Sakakibara, Tatsuo Nakayama, Masahito Suiko, Ken Yanagisawa, Yasunari Takami, and Ming-Cheh Liu
- Subjects
Sulfotransferase ,Molecular Sequence Data ,Biophysics ,Molecular cloning ,Biology ,Transfection ,Biochemistry ,law.invention ,Substrate Specificity ,Evolution, Molecular ,chemistry.chemical_compound ,Mice ,law ,SULT2B1 ,Animals ,Northern blot ,Amino Acid Sequence ,RNA, Messenger ,Cloning, Molecular ,Molecular Biology ,chemistry.chemical_classification ,Sequence Homology, Amino Acid ,Cell Biology ,Sequence Analysis, DNA ,Molecular biology ,Arylsulfotransferase ,Recombinant Proteins ,Amino acid ,Enzyme ,chemistry ,COS Cells ,Recombinant DNA ,Hydroxysteroid ,Sulfotransferases ,Microsatellite Repeats - Abstract
Nucleotide sequences of two mouse cDNAs encoding new sulfotransferase enzymes were determined. Analysis of the deduced amino acid sequences revealed that one represents a novel member of the phenol sulfotransferase family and the other is highly homologous to human SULT2B1 hydroxysteroid sulfotransferases. The recombinant enzymes, transiently expressed in COS-7 cells, were characterized with respect to their substrate specificity using a variety of substrates for different types of sulfotransferases. The tissue-specific expression of these two new mouse sulfotransferases was examined by Northern blot analysis.
- Published
- 1998
49. Enzymology of human cytosolic sulfotransferases
- Author
-
Charles N. Falany
- Subjects
Sulfotransferase ,Chemistry ,food and beverages ,Biological activity ,Sulfotransferase 1A1 ,Biochemistry ,Xenobiotics ,chemistry.chemical_compound ,Sulfation ,Cytosol ,Pharmaceutical Preparations ,SULT2B1 ,Genetics ,Humans ,Estrogen Sulfotransferase ,Hydroxysteroid ,Sulfonic Acids ,Sulfotransferases ,Molecular Biology ,Drug metabolism ,Biotransformation ,Biotechnology - Abstract
Conjugation of many xenobiotics, drugs, and endogenous compounds with a sulfonate moiety is an important reaction in their biotransformation. Sulfation of these compounds generally results in a decrease in biological activity and an increase in their urinary excretion. However, in certain instances, sulfation results in bioactivation to reactive electrophilic or therapeutically active forms. At least four cytosolic sulfotransferases (STs) have been identified and characterized from human tissues. These enzymes are two forms of phenol ST (PST), the phenol-sulfating and the monoamine-sulfating forms of PST (P-PST and M-PST, respectively), an estrogen sulfotransferase (EST), and a hydroxysteroid ST, dehydroepiandrosterone ST (DHEA-ST). Although four cytosolic STs have been well characterized in human tissues, evidence is accumulating for the presence of allelic forms or additional distinct forms of the STs in human tissues. The STs possess distinct but overlapping substrate specificities, and all of the STs are capable of conjugating both xenobiotic and endogenous compounds. The individual forms of ST may display distinct patterns of tissue specific expression and different mechanisms of regulation. Although the role of sulfation in drug metabolism is well recognized, an increased understanding of the biochemistry and molecular biology of the STs should also provide additional information as to their functions in many normal physiologic processes.
- Published
- 1997
50. Sulfation of vitamin D 3 -related compounds-identification and characterization of the responsible human cytosolic sulfotransferases.
- Author
-
Kurogi K, Sakakibara Y, Suiko M, and Liu MC
- Subjects
- Humans, Hydrogen-Ion Concentration, Kinetics, Calcifediol metabolism, Cytosol enzymology, Dehydrocholesterols metabolism, Sulfates metabolism, Sulfotransferases metabolism
- Abstract
While 25-hydroxyvitamin D
3 3-O-sulfate is known to be present in circulation, how it is generated in the body remains unclear. This study aimed to investigate its sulfation in major human organs and to unveil the responsible cytosolic sulfotransferases (SULTs). Of the vitamin D3 -related compounds tested, 25-hydroxyvitamin D3 and 7-dehydrocholesterol are preferentially sulfated by human organ cytosols. Among the 13 human SULTs, SULT2A1 shows sulfating activity toward all vitamin D3 -related compounds, whereas SULT1A1 and SULT2B1a/SULT2B1b show sulfating activity exclusively for, respectively, calcitriol and 7-dehydrocholesterol. These findings suggest that the metabolic pathway leading to the formation of 25-hydroxyvitamin D3 3-O-sulfate may be mediated by the sulfation of 25-hydroxyvitamin D3 or by the conversion of 7-dehydrocholesterol-3-O-sulfate in the skin., (© 2017 Federation of European Biochemical Societies.)- Published
- 2017
- Full Text
- View/download PDF
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