6,863 results on '"Sp1 transcription factor"'
Search Results
2. Treponema denticola dentilisin triggered TLR2/MyD88 activation upregulates a tissue destructive program involving MMPs via Sp1 in human oral cells
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Ganther, Sean, Radaic, Allan, Malone, Erin, Kamarajan, Pachiyappan, Chang, Nai-Yuan Nicholas, Tafolla, Christian, Zhan, Ling, Fenno, J Christopher, and Kapila, Yvonne L
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Biomedical and Clinical Sciences ,Dentistry ,Infectious Diseases ,Dental/Oral and Craniofacial Disease ,Aetiology ,2.1 Biological and endogenous factors ,Inflammatory and immune system ,Infection ,Bacterial Proteins ,Cells ,Cultured ,Humans ,Matrix Metalloproteinases ,Myeloid Differentiation Factor 88 ,Peptide Hydrolases ,Periodontal Diseases ,Periodontal Ligament ,Sp1 Transcription Factor ,Toll-Like Receptor 2 ,Treponema denticola ,Treponemal Infections ,Up-Regulation ,Virulence Factors ,Microbiology ,Immunology ,Medical Microbiology ,Virology ,Medical microbiology - Abstract
Periodontal disease is driven by dysbiosis in the oral microbiome, resulting in over-representation of species that induce the release of pro-inflammatory cytokines, chemokines, and tissue-remodeling matrix metalloproteinases (MMPs) in the periodontium. These chronic tissue-destructive inflammatory responses result in gradual loss of tooth-supporting alveolar bone. The oral spirochete Treponema denticola, is consistently found at significantly elevated levels in periodontal lesions. Host-expressed Toll-Like Receptor 2 (TLR2) senses a variety of bacterial ligands, including acylated lipopolysaccharides and lipoproteins. T. denticola dentilisin, a surface-expressed protease complex comprised of three lipoproteins has been implicated as a virulence factor in periodontal disease, primarily due to its proteolytic activity. While the role of acylated bacterial components in induction of inflammation is well-studied, little attention has been given to the potential role of the acylated nature of dentilisin. The purpose of this study was to test the hypothesis that T. denticola dentilisin activates a TLR2-dependent mechanism, leading to upregulation of tissue-destructive genes in periodontal tissue. RNA-sequencing of periodontal ligament cells challenged with T. denticola bacteria revealed significant upregulation of genes associated with extracellular matrix organization and degradation including potentially tissue-specific inducible MMPs that may play novel roles in modulating host immune responses that have yet to be characterized within the context of oral disease. The Gram-negative oral commensal, Veillonella parvula, failed to upregulate these same MMPs. Dentilisin-induced upregulation of MMPs was mediated via TLR2 and MyD88 activation, since knockdown of expression of either abrogated these effects. Challenge with purified dentilisin upregulated the same MMPs while a dentilisin-deficient T. denticola mutant had no effect. Finally, T. denticola-mediated activation of TLR2/MyD88 lead to the nuclear translocation of the transcription factor Sp1, which was shown to be a critical regulator of all T. denticola-dependent MMP expression. Taken together, these data suggest that T. denticola dentilisin stimulates tissue-destructive cellular processes in a TLR2/MyD88/Sp1-dependent fashion.
- Published
- 2021
3. Melatonin receptor 1A (MTNR1A) gene linkage and association to type 2 diabetes in Italian families.
- Author
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AMIN, M. and GRAGNOLI, C.
- Abstract
OBJECTIVE: Melatonin regulates the mammalian circadian rhythm and plays metabolic functions such as glucose homeostasis. Both melatonin receptors (MTNR1A and MTNR1B, encoded by the MTNR1A and MTNR1B genes, respectively) are expressed in pancreatic beta cells and mediate the glucometabolic roles of melatonin as well as insulin secretion. The MTNR1B gene is a well-known genetic risk factor in type 2 diabetes (T2D); however, little is known about the involvement of the MTNR1A gene in here T2D. We aimed to investigate whether MTNR1A is linked to and/or associated with familial T2D. SUBJECTS AND METHODS: We genotyped 14 single nucleotide polymorphisms within the MTNR1A gene in 212 peninsular Italian families with T2D. We performed parametric linkage and linkage disequilibrium analyses to investigate the role of MTNR1A variants in conferring T2D risk. We considered variants statistically significant if conferring linkage or linkage disequilibrium with p < 0.05. RESULTS: We found 3 novel variants (rs62350392, rs2119883, and rs13147179) significantly linked to and/or associated with T2D in multigenerational Italian families. CONCLUSIONS: This is the first study to report MTNR1A as a novel risk gene in T2D. Functional studies are needed to confirm these results. [ABSTRACT FROM AUTHOR]
- Published
- 2023
4. Matrix stiffness induces a tumorigenic phenotype in mammary epithelium through changes in chromatin accessibility.
- Author
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Stowers, Ryan S, Shcherbina, Anna, Israeli, Johnny, Gruber, Joshua J, Chang, Julie, Nam, Sungmin, Rabiee, Atefeh, Teruel, Mary N, Snyder, Michael P, Kundaje, Anshul, and Chaudhuri, Ovijit
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Epithelium ,Cell Line ,Tumor ,Extracellular Matrix ,Chromatin ,Epithelial Cells ,Humans ,Breast Neoplasms ,Transcription Factors ,Cell Culture Techniques ,Mechanotransduction ,Cellular ,Phenotype ,Female ,Sp1 Transcription Factor ,Tumor Microenvironment ,Cancer ,Human Genome ,Genetics ,Breast Cancer - Abstract
In breast cancer, the increased stiffness of the extracellular matrix is a key driver of malignancy. Yet little is known about the epigenomic changes that underlie the tumorigenic impact of extracellular matrix mechanics. Here, we show in a three-dimensional culture model of breast cancer that stiff extracellular matrix induces a tumorigenic phenotype through changes in chromatin state. We found that increased stiffness yielded cells with more wrinkled nuclei and with increased lamina-associated chromatin, that cells cultured in stiff matrices displayed more accessible chromatin sites, which exhibited footprints of Sp1 binding, and that this transcription factor acts along with the histone deacetylases 3 and 8 to regulate the induction of stiffness-mediated tumorigenicity. Just as cell culture on soft environments or in them rather than on tissue-culture plastic better recapitulates the acinar morphology observed in mammary epithelium in vivo, mammary epithelial cells cultured on soft microenvironments or in them also more closely replicate the in vivo chromatin state. Our results emphasize the importance of culture conditions for epigenomic studies, and reveal that chromatin state is a critical mediator of mechanotransduction.
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- 2019
5. miR‐138‐5p inhibits the progression of colorectal cancer via regulating SP1/LGR5 axis.
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Zou, Qing‐Tao, Lin, Yi, and Luo, Qing‐Yu
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REVERSE transcriptase polymerase chain reaction , *COLORECTAL cancer , *G protein coupled receptors , *CANCER invasiveness - Abstract
Colorectal cancer (CRC) is one of the main malignancies that seriously threaten human health. Considering the high mortality and morbidity associated with this disease, even surgical resection and chemotherapy may not be sufficient in certain cases. This study aimed to explore the molecular mechanisms of miR‐138‐5p in regulating CRC progression. Quantitative reverse transcriptase polymerase chain reaction and western blot were performed to assess the levels of mRNA and proteins, including miR‐138‐5p, leucine‐rich repeat‐containing G protein‐coupled receptor 5 (LGR5), SP1, β‐catenin, cyclin D1, and c‐myc. The bioactivities of LoVo and HCT116 cells were assessed via MTT assay, flow cytometry, and transwell assay. StarBase was used to identify the downstream targets of genes. Double luciferase reporter and RIP assays revealed the direct binding of miR‐138‐5p to SP1 and of SP1 to LGR5. Our results illustrated that miR‐138‐5p was downregulated in CRC and its knockdown accelerated CRC progression. Conversely, SP1 was upregulated in CRC and its knockdown inhibited CRC progression. SP1 is also targeted by miR‐138‐5p and binds to LGR5. This study showed that miR‐138‐5p inhibits LoVo and HCT116 cell proliferation, migration, and invasion. Overall, miR‐138‐5p regulates CRC progression and promotes apoptosis via the SP1/LGR5 axis. This study indicates that miR‐138‐5p is involved in regulating CRC progression. [ABSTRACT FROM AUTHOR]
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- 2023
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6. Selenium Drives a Transcriptional Adaptive Program to Block Ferroptosis and Treat Stroke.
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Alim, Ishraq, Caulfield, Joseph T, Chen, Yingxin, Swarup, Vivek, Geschwind, Daniel H, Ivanova, Elena, Seravalli, Javier, Ai, Youxi, Sansing, Lauren H, Ste Marie, Emma J, Hondal, Robert J, Mukherjee, Sushmita, Cave, John W, Sagdullaev, Botir T, Karuppagounder, Saravanan S, and Ratan, Rajiv R
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Neurons ,Animals ,Humans ,Mice ,Brain Ischemia ,Intracranial Hemorrhages ,Disease Models ,Animal ,Selenium ,Transcription ,Genetic ,Gene Expression Regulation ,Enzymologic ,Male ,Sp1 Transcription Factor ,Transcription Factor AP-2 ,Stroke ,Cell-Penetrating Peptides ,Endoplasmic Reticulum Stress ,Ferroptosis ,Phospholipid Hydroperoxide Glutathione Peroxidase ,GPX4 ,adaptation ,cell death ,ferroptosis ,intracerebral hemorrhage ,selenium ,selenoprotein ,stroke ,therapeutic peptides ,transcription ,Disease Models ,Animal ,Transcription ,Genetic ,Gene Expression Regulation ,Enzymologic ,Complementary and Alternative Medicine ,Genetics ,Brain Disorders ,Nutrition ,Neurosciences ,1.1 Normal biological development and functioning ,Biological Sciences ,Medical and Health Sciences ,Developmental Biology - Abstract
Ferroptosis, a non-apoptotic form of programmed cell death, is triggered by oxidative stress in cancer, heat stress in plants, and hemorrhagic stroke. A homeostatic transcriptional response to ferroptotic stimuli is unknown. We show that neurons respond to ferroptotic stimuli by induction of selenoproteins, including antioxidant glutathione peroxidase 4 (GPX4). Pharmacological selenium (Se) augments GPX4 and other genes in this transcriptional program, the selenome, via coordinated activation of the transcription factors TFAP2c and Sp1 to protect neurons. Remarkably, a single dose of Se delivered into the brain drives antioxidant GPX4 expression, protects neurons, and improves behavior in a hemorrhagic stroke model. Altogether, we show that pharmacological Se supplementation effectively inhibits GPX4-dependent ferroptotic death as well as cell death induced by excitotoxicity or ER stress, which are GPX4 independent. Systemic administration of a brain-penetrant selenopeptide activates homeostatic transcription to inhibit cell death and improves function when delivered after hemorrhagic or ischemic stroke.
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- 2019
7. Sp1-Induced SETDB1 Overexpression Transcriptionally Inhibits HPGD in a β-Catenin-Dependent Manner and Promotes the Proliferation and Metastasis of Gastric Cancer.
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Yaguan Fan, Libo Yang, Yi Ren, Yunhua Wu, Linhai Li, and Lihua Li
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REVERSE transcriptase polymerase chain reaction , *STOMACH cancer , *HEMATOXYLIN & eosin staining , *GENETIC overexpression , *WESTERN immunoblotting - Abstract
Purpose: Gastric cancer (GC) has high morbidity and mortality, the cure rate of surgical treatment and drug chemotherapy is not ideal. Therefore, development of new treatment strategies is necessary. We aimed to identify the mechanism underlying Sp1 regulation of GC progression. Methods and Methods: The levels of Sp1, β-catenin, SET domain bifurcated 1 (SETDB1), and 15-hydroxyprostaglandin dehydrogenase (HPGD) were detected by quantitative reverse transcription polymerase chain reaction and western blot analysis. The targets of SETDB1 were predicted by AnimalTFDB, and dual-luciferase reporter assay was used for confirming the combination of Sp1, β-catenin, and SETDB1. HGC27 or AGS cells (1×106 cells/mouse) were injected into mice via the caudal vein for GC model establishment. The level of Ki67 was detected using immunohistochemistry, and hematoxylin and eosin staining was performed for evaluating tumor metastasis in mice with GC. Results: HPGD was inhibited, while the protein levels of Sp1, β-catenin, and SETDB1 were up-regulated in GC tissues and cell lines. HPGD overexpression or SETDB1 silencing inhibited the proliferation, invasion, and migration of GC cells, and Sp1 regulated the proliferation, invasion, and migration of GC cells in a β-catenin-dependent manner. Furthermore, HPGD served as a target of SETDB1, and it was negatively regulated by SETDB1; additionally, Sp1 and β-catenin bound to the SETDB1 promoter and negatively regulated HPGD expression. We proved that Sp1 regulated GC progression via the SETDB1/HPGD axis. Conclusions: Our findings revealed that Sp1 transcriptionally inhibited HPGD via SETDB1 in a β-catenin-dependent manner and promoted the proliferation and metastasis of GC cells. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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8. Mithramycin and its analogs: Molecular features and antitumor action.
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Portugal, José
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TRANSCRIPTION factor Sp1 , *TRANSCRIPTION factors , *ANTINEOPLASTIC antibiotics , *EWING'S sarcoma , *GENE expression - Abstract
The antitumor antibiotic mithramycin A (MTA) binds to G/C-rich DNA sequences in the presence of dications. MTA inhibits transcription regulated by the Sp1 transcription factor, often enhanced during tumor development. It shows antitumor activity, but its clinical use was discontinued due to toxic side effects. However, recent observations have led to its use being reconsidered. The MTA biosynthetic pathways have been modified to produce mithramycin analogs (mithralogs) that encompass lower toxicity and improved pharmacological activity. Some mithralogs reduce gene expression in human ovarian and prostate tumors, among other types of cancer. They down-regulate gene expression in various cellular processes, including Sp1-responsive genes that control tumor development. Moreover, MTA and several mithralogs, such as EC-8042 (DIG-MSK) and EC-8105, effectively treat Ewing sarcoma by inhibiting transcription controlled by the oncogenic EWS-FLI1 transcription factor. [ABSTRACT FROM AUTHOR]
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- 2024
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9. RNA-binding protein Rbpms is represented in human retinas by isoforms A and C and its transcriptional regulation involves Sp1-binding site
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Ye, Linda, Gu, Lei, Caprioli, Joseph, and Piri, Natik
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Biochemistry and Cell Biology ,Bioinformatics and Computational Biology ,Biological Sciences ,Genetics ,Underpinning research ,1.1 Normal biological development and functioning ,Eye Proteins ,HEK293 Cells ,Humans ,Mutagenesis ,Site-Directed ,Mutation ,NF-kappa B ,Protein Isoforms ,RNA-Binding Proteins ,Response Elements ,Retina ,Sp1 Transcription Factor ,Transcription ,Genetic ,Retinal ganglion cells ,Transcription factors ,CpG islands ,Promoter ,Plant Biology ,Genetics & Heredity ,Plant Biology & Botany ,Plant biology - Abstract
Rbpms (RNA-binding protein with multiple splicing) is a member of the RRM (RNA Recognition Motif) family of RNA-binding proteins, which is expressed as multiple alternatively spliced transcripts encoding different protein isoforms. We have shown earlier that Rbpms expression in the retina is restricted to retinal ganglion cells (RGCs), and have characterized this gene as a marker for RGCs. The aim of this study was to identify isoforms representing Rbpms in human retinas and to analyze its transcriptional regulation. We found that Rbpms is expressed as transcription variants 1 and 3 encoding isoforms A and C, respectively. These isoforms are encoded by the same first 6 exons but have different C-terminal ends encoded by exon 8 in variant 1 and exon 7 in variant 3. Computational analysis of the Rbpms 5' untranslated and flanking regions reveals the presence of three CpG islands and four predicted promoter regions (PPRs). The effect of PPR 1 (- 1672/- 1420) and PPR2 (- 330/- 79) on transcriptional activation was minimal, whereas PPR 3 (- 73/+ 177) and PPR4 (+ 274/+ 524) induced the expression by ~ 7 and ninefold compared to control, respectively. The maximum activity, a 30-fold increase above the control level, was obtained from the construct containing both PPRs 3 and 4. Site-directed mutagenesis of several cis-elements within PPR3 and PPR4 including five for Sp1, one for AP1, and two for NF-kB showed that mutation of the first three and especially the first GC box resulted in a threefold downregulation of gene expression. AP1, NF-kB, and two downstream Sp1 sites had no significant effect on expression level. The possible involvement of the GC box 1 at position - 54 in transcriptional regulation of Rbpms was corroborated by EMSA, which showed formation of a DNA-protein complex in the presence of the oligonucleotide corresponding to this Sp1-binding site.
- Published
- 2018
10. BCI, an inhibitor of the DUSP1 and DUSP6 dual specificity phosphatases, enhances P2X7 receptor expression in neuroblastoma cells
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María Benito-León, Juan Carlos Gil-Redondo, Raquel Perez-Sen, Esmerilda G. Delicado, Felipe Ortega, and Rosa Gomez-Villafuertes
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P2X7 receptor ,neuroblastoma ,DUSP1 ,p38 phosphorylation ,Sp1 transcription factor ,phosphatase inhibitor ,Biology (General) ,QH301-705.5 - Abstract
P2X7 receptor (P2RX7) is expressed strongly by most human cancers, including neuroblastoma, where high levels of P2RX7 are correlated with a poor prognosis for patients. Tonic activation of P2X7 receptor favors cell metabolism and angiogenesis, thereby promoting cancer cell proliferation, immunosuppression, and metastasis. Although understanding the mechanisms that control P2X7 receptor levels in neuroblastoma cells could be biologically and clinically relevant, the intracellular signaling pathways involved in this regulation remain poorly understood. Here we show that (E)-2-benzylidene-3-(cyclohexylamino)-2,3-dihydro-1H-inden-1-one (BCI), an allosteric inhibitor of dual specificity phosphatases (DUSP) 1 and 6, enhances the expression of P2X7 receptor in N2a neuroblastoma cells. We found that exposure to BCI induces the phosphorylation of mitogen-activated protein kinases p38 and JNK, while it prevents the phosphorylation of ERK1/2. BCI enhanced dual specificity phosphatase 1 expression, whereas it induced a decrease in the dual specificity phosphatase 6 transcripts, suggesting that BCI-dependent inhibition of dual specificity phosphatase 1 may be responsible for the increase in p38 and JNK phosphorylation. The weaker ERK phosphorylation induced by BCI was reversed by p38 inhibition, indicating that this MAPK is involved in the regulatory loop that dampens ERK activity. The PP2A phosphatase appears to be implicated in the p38-dependent dephosphorylation of ERK1/2. In addition, the PTEN phosphatase inhibition also prevented ERK1/2 dephosphorylation, probably through p38 downregulation. By contrast, inhibition of the p53 nuclear factor decreased ERK phosphorylation, probably enhancing the activity of p38. Finally, the inhibition of either p38 or Sp1-dependent transcription halved the increase in P2X7 receptor expression induced by BCI. Moreover, the combined inhibition of both p38 and Sp1 completely prevented the effect exerted by BCI. Together, our results indicate that dual specificity phosphatase 1 acts as a novel negative regulator of P2X7 receptor expression in neuroblastoma cells due to the downregulation of the p38 pathway.
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- 2022
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11. Lyon IARC Polyomavirus Displays Transforming Activities in Primary Human Cells.
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Venuti, Assunta, Carmen Romero-Medina, Maria, Melita, Giusi, Grazia Ceraolo, Maria, Brancaccio, Rosario Nicola, Sirand, Cecilia, Taverniti, Valerio, Steenbergen, Renske, Gheit, Tarik, and Tommasino, Massimo
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TELOMERASE reverse transcriptase , *TRANSCRIPTION factor Sp1 , *POLYOMAVIRUSES , *VETERINARY virology , *MERKEL cells ,KERATINOCYTE differentiation - Abstract
Several studies reported the presence of a recently discovered polyomavirus (PyV), Lyon IARC PyV (LIPyV), in human and domestic animal specimens. LIPyV has some structural similarities to well-established animal and human oncogenic PyVs, such as raccoon PyV and Merkel cell PyV (MCPyV), respectively. In this study, we demonstrate that LIPyV early proteins immortalize human foreskin keratinocytes. LIPyV LT binds pRb, accordingly cell cycle checkpoints are altered in primary human fibroblasts and keratinocytes expressing LIPyV early genes. Mutation of the pRb binding site in LT strongly affected the ability of LIPyV ER to induced HFK immortalization. LIPyV LT also binds p53 and alters p53 functions activated by cellular stresses. Finally, LIPyV early proteins activate telomerase reverse transcriptase (hTERT) gene expression, via accumulation of the Sp1 transcription factor. Sp1 recruitment to the hTERT promoter is controlled by its phosphorylation, which is mediated by ERK1 and CDK2. Together, these data highlight the transforming properties of LIPyV in in vitro experimental models, supporting its possible oncogenic nature. IMPORTANCE Lyon IARC PyV is a recently discovered polyomavirus that shows some structural similarities to well-established animal and human oncogenic PyVs, such as raccoon PyV and Merkel cell PyV, respectively. Here, we show the capability of LIPyV to efficiently promote cellular transformation of primary human cells, suggesting a possible oncogenic role of this virus in domestic animals and/or humans. Our study identified a novel virus-mediated mechanism of activation of telomerase reverse transcriptase gene expression, via accumulation of the Sp1 transcription factor. In addition, because the persistence of infection is a key event in virus-mediated carcinogenesis, it will be important to determine whether LIPyV can deregulate immunerelated pathways, similarly to the well-established oncogenic viruses. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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12. A multicenter, phase 1, Adult Brain Tumor Consortium trial of oral terameprocol for patients with recurrent high-grade glioma (GATOR).
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Ahluwalia MS, Ozair A, Rudek M, Ye X, Holdhoff M, Lieberman FS, Piotrowski AF, Nabors B, Desai A, Lesser G, Huang RC, Glenn S, Khosla AA, Peereboom DM, Wen PY, and Grossman SA
- Subjects
- Humans, Male, Middle Aged, Adult, Female, Aged, Administration, Oral, Aged, 80 and over, Neoplasm Recurrence, Local drug therapy, Neoplasm Recurrence, Local pathology, Neoplasm Grading, Maximum Tolerated Dose, Glioma drug therapy, Glioma pathology, Brain Neoplasms drug therapy, Brain Neoplasms pathology
- Abstract
Recurrent high-grade gliomas (rHGGs) have a dismal prognosis, where the maximum tolerated dose (MTD) of IV terameprocol (5 days/month), a transcriptional inhibitor of specificity protein 1 (Sp1)-regulated proteins, is 1,700 mg/day with median area under the plasma concentration-time curve (AUC) of 31.3 μg∗h/mL. Given potentially increased efficacy with sustained systemic exposure and challenging logistics of daily IV therapy, here we investigate oral terameprocol for rHGGs in a multicenter, phase 1 trial (GATOR). Using a 3 + 3 dose-escalation design, we enroll 20 patients, with median age 60 years (range 31-80), 70% male, and median one relapse (range 1-3). Fasting patients tolerate 1,200 mg/day (n = 3), 2,400 mg/day (n = 6), 3,600 mg/day (n = 3), and 6,000 mg/day (n = 2) oral doses without major toxicities. However, increased dosage does not lead to increased systemic exposure, including in fed state (6,000 mg/day, n = 4), with maximal AUC <5 μg∗h/mL. These findings warrant trials investigating approaches that provide sustained systemic levels of transcription inhibitors to exploit their therapeutic potential. This study was registered at ClinicalTrials.gov (NCT02575794)., Competing Interests: Declaration of interests M.S.A. – grants: Seagen, AstraZeneca, BMS, Bayer, Incyte, Pharmacyclics, Novocure, MimiVax, and Merck. Consultation fees: Bayer, Novocure, Kiyatec, Insightec, GSK, Xoft, Nuvation, Celularity, SDP Oncology, Apollomics, Prelude Therapeutics, Janssen, Tocagen, Voyager Therapeutics, ViewRay, Caris Life Sciences, Pyramid Biosciences, Varian Medical Systems, Cairn Therapeutics, AnHeart Therapeutics, Menarini Ricerche, Sumitomo Pharma Oncology, Autem therapeutics, GT Medical Technologies, Allovir, Equillium Bio., QV Bioelectronics, and Theraguix. Scientific Advisory Board memberships: Cairn Therapeutics, Pyramid Biosciences, Bugworks, and Modifi Biosciences. Data Safety and Monitoring Committee membership: VBI Vaccines. Stock shareholder: MimiVax, CytoDyn, Trisalus Lifesciences, and MedInnovate Advisors, LLC. D.M.P. – consulting or advisory role: Orbus Therapeutics, Sumitomo Dainippon Pharma Oncology, Inc., Stemline Therapeutics, and Novocure. Research funding: Pfizer (Inst), Novartis (Inst), NeOnc Technologies (Inst), Orbus Therapeutics (Inst), Bristol Myers Squibb (Inst), Genentech/Roche (Inst), Pharmacyclics (Inst), Bayer (Inst), Karyopharm Therapeutics (Inst), Apollomics (Inst), Vigeo Therapeutics (Inst), Global Coalition for Adaptive Research (Inst), MimiVax (Inst), Ono Pharmaceutical (Inst), and Mylan (Inst). Equity ownership/stock options: Pfizer (Pharmaceuticals) and Gilead (Pharmaceuticals). M.H. – Data Safety Monitoring Board member: Parexel and Advarra. Institutional research funding: Novartis and Vanquish. P.Y.W. – research support: AstraZeneca, Black Diamond, Bristol Meyers Squibb, Celgene, Chimerix, Eli Lily, Erasca, Genentech/Roche, Kazia, MediciNova, Merck, Novartis, Nuvation Bio, Servier, Vascular Biogenics, and VBI Vaccines. Advisory board/consultant: AstraZeneca, Black Diamond, Celularity, Chimerix, Day One Bio, Genenta, Glaxo Smith Kline, Merck, Mundipharma, Novartis, Novocure, Nuvation Bio, Prelude Therapeutics, Sapience, Servier, Sagimet, Vascular Biogenics, and VBI Vaccines. R.C.H. has patents related to the drug terameprocol and is affiliated with Erimos Pharmaceuticals, which holds rights to the drug. S.G. is an independent contractor/consultant to Erimos Pharmaceuticals, LLC with no ownership or other financial interest in Erimos Pharmaceuticals or its products., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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13. The WNK1 kinase regulates the stability of transcription factors during wound healing of human corneal epithelial cells.
- Author
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Desjardins, Pascale, Le‐Bel, Gaëtan, Ghio, Sergio C., Germain, Lucie, and Guérin, Sylvain L.
- Subjects
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TRANSCRIPTION factors , *WOUND healing , *AP-1 transcription factor , *EPITHELIAL cells , *CORNEA , *UBIQUITINATION - Abstract
Due to its superficial anatomical localization, the cornea is continuously subjected to injuries. Damages to the corneal epithelium trigger important changes in the composition of the extracellular matrix to which the basal human corneal epithelial cells (hCECs) attach. These changes are perceived by membrane‐bound integrins and ultimately lead to re‐epithelialization of the injured epithelium through intracellular signalin. Among the many downstream targets of the integrin‐activated signaling pathways, WNK1 is the kinase whose activity is the most strongly increased during corneal wound healing. We previously demonstrated that pharmacological inhibition of WNK1 prevents proper closure of wounded human tissue‐engineered cornea in vitro. In the present study, we investigated the molecular mechanisms by which WNK1 contributes to corneal wound healing. By exploiting transcription factors microarrays, electrophoretic mobility‐shift assay, and gene profiling analyses, we demonstrated that the DNA binding properties and expression of numerous transcription factors (TFs), including the well‐known, ubiquitous TFs specific protein 1 (Sp1) and activator protein 1 (AP1), were reduced in hCECs upon WNK1 inhibition by WNK463. This process appears to be mediated at least in part by alteration in both the ubiquitination and glycosylation status of these TFs. These changes in TFs activity and expression impacted the transcription of several genes, including that encoding the α5 integrin subunit, a well‐known target of both Sp1 and AP1. Gene profiling revealed that only a moderate number of genes in hCECs had their level of expression significantly altered in response to WNK463 exposition. Interestingly, analysis of the microarray data for these deregulated genes using the ingenuity pathway analysis software predicted that hCECs would stop migrating and proliferating but differentiate more when they are grown in the presence of the WNK1 inhibitor. These results demonstrate that WNK1 plays a critical function by orienting hCECs into the appropriate biological response during the process of corneal wound healing. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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14. Cardiac myocyte p38α kinase regulates angiogenesis via myocyte-endothelial cell cross-talk during stress-induced remodeling in the heart
- Author
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Rose, Beth A, Yokota, Tomohiro, Chintalgattu, Vishnu, Ren, Shuxun, Iruela-Arispe, Luisa, Khakoo, Aarif Y, Minamisawa, Susumu, and Wang, Yibin
- Subjects
Biochemistry and Cell Biology ,Medical Physiology ,Biomedical and Clinical Sciences ,Biological Sciences ,Heart Disease ,Heart Disease - Coronary Heart Disease ,Cardiovascular ,Aetiology ,2.1 Biological and endogenous factors ,Animals ,Animals ,Newborn ,Cell Hypoxia ,Cells ,Cultured ,Crosses ,Genetic ,Endothelium ,Vascular ,Enzyme Activation ,Gene Expression Regulation ,Human Umbilical Vein Endothelial Cells ,Humans ,Male ,Mice ,Knockout ,Mice ,Transgenic ,Mitogen-Activated Protein Kinase 14 ,Myocardial Ischemia ,Myocardial Revascularization ,Myocytes ,Cardiac ,RNA Interference ,Rats ,Sprague-Dawley ,Recombinant Proteins ,Sp1 Transcription Factor ,Sus scrofa ,Vascular Endothelial Growth Factor A ,Vascular Endothelial Growth Factor Receptor-2 ,angiogenesis ,cardiac hypertrophy ,cardiomyocyte ,p38 MAPK ,vascular endothelial growth factor ,cross-talk ,Chemical Sciences ,Medical and Health Sciences ,Biochemistry & Molecular Biology ,Biological sciences ,Biomedical and clinical sciences ,Chemical sciences - Abstract
Stress-induced p38 mitogen-activated protein kinase (MAPK) activity is implicated in pathological remodeling in the heart. For example, constitutive p38 MAPK activation in cardiomyocytes induces pathological features, including myocyte hypertrophy, apoptosis, contractile dysfunction, and fetal gene expression. However, the physiological function of cardiomyocyte p38 MAPK activity in beneficial compensatory vascular remodeling is unclear. This report investigated the functional role and the underlying mechanisms of cardiomyocyte p38 MAPK activity in cardiac remodeling induced by chronic stress. Using both in vitro and in vivo model systems, we found that p38 MAPK activity is required for hypoxia-induced pro-angiogenic activity from cardiomyocytes and that p38 MAPK activation in cardiomyocyte is sufficient to promote paracrine signaling-mediated, pro-angiogenic activity. We further demonstrate that VEGF is a paracrine factor responsible for the p38 MAPK-mediated pro-angiogenic activity from cardiomyocytes and that p38 MAPK pathway activation is sufficient for inducing VEGF secretion from cardiomyocytes in an Sp1-dependent manner. More significantly, cardiomyocyte-specific inactivation of p38α in mouse heart impaired compensatory angiogenesis after pressure overload and promoted early onset of heart failure. In summary, p38αMAPK has a critical role in the cross-talk between cardiomyocytes and vasculature by regulating stress-induced VEGF expression and secretion in cardiomyocytes. We conclude that as part of a stress-induced signaling pathway, p38 MAPK activity significantly contributes to both pathological and compensatory remodeling in the heart.
- Published
- 2017
15. Discovery of Novel Human Gene Regulatory Modules from Gene Co-expression and Promoter Motif Analysis.
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Ma, Shisong, Snyder, Michael, and Dinesh-Kumar, Savithramma P
- Subjects
Humans ,Lysine ,CCAAT-Binding Factor ,Oligonucleotide Array Sequence Analysis ,Gene Expression Regulation ,Epigenesis ,Genetic ,Methylation ,Sp1 Transcription Factor ,Interferon Regulatory Factor-1 ,YY1 Transcription Factor ,Gene Regulatory Networks ,Promoter Regions ,Genetic ,Nucleotide Motifs ,Regulatory Factor X1 ,Genetics ,Human Genome ,Biotechnology ,2.1 Biological and endogenous factors ,Generic Health Relevance ,Epigenesis ,Genetic ,Promoter Regions ,Biochemistry and Cell Biology ,Other Physical Sciences - Abstract
Deciphering gene regulatory networks requires identification of gene expression modules. We describe a novel bottom-up approach to identify gene modules regulated by cis-regulatory motifs from a human gene co-expression network. Target genes of a cis-regulatory motif were identified from the network via the motif's enrichment or biased distribution towards transcription start sites in the promoters of co-expressed genes. A gene sub-network containing the target genes was extracted and used to derive gene modules. The analysis revealed known and novel gene modules regulated by the NF-Y motif. The binding of NF-Y proteins to these modules' gene promoters were verified using ENCODE ChIP-Seq data. The analyses also identified 8,048 Sp1 motif target genes, interestingly many of which were not detected by ENCODE ChIP-Seq. These target genes assemble into house-keeping, tissues-specific developmental, and immune response modules. Integration of Sp1 modules with genomic and epigenomic data indicates epigenetic control of Sp1 targets' expression in a cell/tissue specific manner. Finally, known and novel target genes and modules regulated by the YY1, RFX1, IRF1, and 34 other motifs were also identified. The study described here provides a valuable resource to understand transcriptional regulation of various human developmental, disease, or immunity pathways.
- Published
- 2017
16. BALR-6 regulates cell growth and cell survival in B-lymphoblastic leukemia.
- Author
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Rodríguez-Malavé, Norma I, Fernando, Thilini R, Patel, Parth C, Contreras, Jorge R, Palanichamy, Jayanth Kumar, Tran, Tiffany M, Anguiano, Jaime, Davoren, Michael J, Alberti, Michael O, Pioli, Kimanh T, Sandoval, Salemiz, Crooks, Gay M, and Rao, Dinesh S
- Subjects
Hematopoietic Stem Cells ,Cell Line ,Tumor ,Animals ,Humans ,Mice ,Cell Proliferation ,Cell Survival ,Sp1 Transcription Factor ,Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ,Gene Knockdown Techniques ,Transcriptome ,RNA ,Long Noncoding ,lncRNA ,B-ALL ,MLL ,SP1 ,Microarray ,Leukemia ,RNA ,Non-coding RNA ,Cell Line ,Tumor ,Long Noncoding ,Oncology & Carcinogenesis ,Oncology and Carcinogenesis - Abstract
BackgroundA new class of non-coding RNAs, known as long non-coding RNAs (lncRNAs), has been recently described. These lncRNAs are implicated to play pivotal roles in various molecular processes, including development and oncogenesis. Gene expression profiling of human B-ALL samples showed differential lncRNA expression in samples with particular cytogenetic abnormalities. One of the most promising lncRNAs identified, designated B-ALL associated long RNA-6 (BALR-6), had the highest expression in patient samples carrying the MLL rearrangement, and is the focus of this study.ResultsHere, we performed a series of experiments to define the function of BALR-6, including several novel splice forms that we identified. Functionally, siRNA-mediated knockdown of BALR-6 in human B-ALL cell lines caused reduced cell proliferation and increased cell death. Conversely, overexpression of BALR-6 isoforms in both human and mouse cell lines caused increased proliferation and decreased apoptosis. Overexpression of BALR-6 in murine bone marrow transplantation experiments caused a significant increase in early hematopoietic progenitor populations, suggesting that its dysregulation may cause developmental changes. Notably, the knockdown of BALR-6 resulted in global dysregulation of gene expression. The gene set was enriched for leukemia-associated genes, as well as for the transcriptome regulated by Specificity Protein 1 (SP1). We confirmed changes in the expression of SP1, as well as its known interactor and downstream target CREB1. Luciferase reporter assays demonstrated an enhancement of SP1-mediated transcription in the presence of BALR-6. These data provide a putative mechanism for regulation by BALR-6 in B-ALL.ConclusionsOur findings support a role for the novel lncRNA BALR-6 in promoting cell survival in B-ALL. Furthermore, this lncRNA influences gene expression in B-ALL in a manner consistent with a function in transcriptional regulation. Specifically, our findings suggest that BALR-6 expression regulates the transcriptome downstream of SP1, and that this may underlie the function of BALR-6 in B-ALL.
- Published
- 2015
17. AMPK-SP1-Guided Dynein Expression Represents a New Energy-Responsive Mechanism and Therapeutic Target for Diabetic Nephropathy.
- Author
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Williquett J, Allamargot C, and Sun H
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- Animals, Humans, AMP-Activated Protein Kinases metabolism, AMP-Activated Protein Kinases genetics, Energy Metabolism drug effects, Sp1 Transcription Factor, Diabetic Nephropathies metabolism, Diabetic Nephropathies genetics, Dyneins metabolism, Dyneins genetics
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- 2024
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18. MicroRNA-199-3p targets Sp1 transcription factor to regulate proliferation and epithelial to mesenchymal transition of human lung cancer cells.
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Fu, Jiajia, Li, Tong, Jiang, Xiaozhen, Xia, Bin, and Hu, Lijuan
- Subjects
- *
TRANSCRIPTION factor Sp1 , *CANCER cells , *EPITHELIAL-mesenchymal transition , *LUNG cancer , *CELL cycle , *METASTASIS - Abstract
The present study was undertaken to study the function of miRNA-199-3p in the regulation of human lung cancer growth and metastasis. The results showed significant (P < 0.05) downregulation of miRNA-199-3p in lung cancer tissues and cell lines. Overexpression of miR-197 caused considerable inhibition of the viability and colony formation of the lung cancer cells. The inhibition of proliferation was found to be due to the arrest of the SK-LU-1 lung cancer cells. At the G2/M phase of the cell cycle. In silico analysis and subsequent the dual-luciferase assays showed that miR-199-3p targets Sp1 at molecular. The expression of Sp1 was significantly (P < 0.05) upregulated in lung cancer cells and tissues. Nonetheless, miR-199-3p overexpression could cause post-transcriptional suppression of Sp1. Silencing of Sp1suppress the proliferation of SK-LU-1 lung cancer cells. However, overexpression Sp1 transcription factor prevents the tumor-suppressive effects of miR-199-3p on lung cancer cells. Additionally, miR-199-3p was found to suppresses the migration, invasion and epithelial-to-mesenchymal transition of human lung cancer cells. Summing up, miRNA-199-3p/SP1 axis controls the growth and metastasis of SK-LU-1 lung cancer cells. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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19. Structure of a DNA G-quadruplex that Modulates SP1 Binding Sites Architecture in HIV-1 Promoter.
- Author
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De Rache, Aurore, Marquevielle, Julien, Bouaziz, Serge, Vialet, Brune, Andreola, Marie-Line, Mergny, Jean-Louis, and Amrane, Samir
- Subjects
- *
QUADRUPLEX nucleic acids , *BINDING sites , *TRANSCRIPTION factor Sp1 , *GENETIC regulation , *HIV , *NUCLEIC acids , *DNA structure - Abstract
[Display omitted] • Context: Nucleic acid sequences containing guanine tracts can form G-quadruplexes (G4s), impacting gene regulation. • Principal results : HIVpro2 DNA sequence, derived from the HIV-1 promoter, forms a hybrid G4 structure with a single-nucleotide bulge. The structural interplay between HIVpro2 and other G4s can modulate SP1 binding sites architecture, potentially regulating viral transcription and latency. • Conceptual advance : Discovery of a G4 structure in HIVpro2 suggests a novel mechanism for HIV-1 gene regulation. • Significance : Understanding the structural switch between G4s and canonical duplexes may offer new strategies for HIV-1 therapy and latency control. Nucleic acid sequences containing guanine tracts are able to form non-canonical DNA or RNA structures known as G-quadruplexes (or G4s). These structures, based on the stacking of G-tetrads, are involved in various biological processes such as gene expression regulation. Here, we investigated a G4 forming sequence, HIVpro2, derived from the HIV-1 promoter. This motif is located 60 nucleotides upstream of the proviral Transcription Starting Site (TSS) and overlaps with two SP1 transcription factor binding sites. Using NMR spectroscopy, we determined that HIVpro2 forms a hybrid type G4 structure with a core that is interrupted by a single nucleotide bulge. An additional reverse-Hoogsteen AT base pair is stacked on top of the tetrad. SP1 transcription factor is known to regulate transcription activity of many genes through the recognition of Guanine-rich duplex motifs. Here, the formation of HIVpro2 G4 may modulate SP1 binding sites architecture by competing with the formation of the canonical duplex structure. Such DNA structural switch potentially participates to the regulation of viral transcription and may also interfere with HIV-1 reactivation or viral latency. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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- View/download PDF
20. Genetic Variants in Transcription Factors Are Associated With the Pharmacokinetics and Pharmacodynamics of Metformin
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Goswami, S, Yee, SW, Stocker, S, Mosley, JD, Kubo, M, Castro, R, Mefford, JA, Wen, C, Liang, X, Witte, J, Brett, C, Maeda, S, Simpson, MD, Hedderson, MM, Davis, RL, Roden, DM, Giacomini, KM, and Savic, RM
- Subjects
Pharmacology and Pharmaceutical Sciences ,Biomedical and Clinical Sciences ,Clinical Sciences ,Diabetes ,Genetics ,Human Genome ,Clinical Research ,Metabolic and endocrine ,Adult ,Aged ,Aged ,80 and over ,Biomarkers ,Diabetes Mellitus ,Type 2 ,Female ,Genome-Wide Association Study ,Glycated Hemoglobin ,Hepatocyte Nuclear Factor 4 ,Homozygote ,Humans ,Hypoglycemic Agents ,Male ,Metformin ,Middle Aged ,Models ,Biological ,Multivariate Analysis ,PPAR alpha ,Pharmacogenetics ,Phenotype ,Polymorphism ,Single Nucleotide ,Retrospective Studies ,Sp1 Transcription Factor ,Transcription Factors ,Treatment Outcome ,United States ,Young Adult ,Pharmacology & Pharmacy ,Pharmacology and pharmaceutical sciences - Abstract
One-third of type 2 diabetes patients do not respond to metformin. Genetic variants in metformin transporters have been extensively studied as a likely contributor to this high failure rate. Here, we investigate, for the first time, the effect of genetic variants in transcription factors on metformin pharmacokinetics (PK) and response. Overall, 546 patients and healthy volunteers contributed their genome-wide, pharmacokinetic (235 subjects), and HbA1c data (440 patients) for this analysis. Five variants in specificity protein 1 (SP1), a transcription factor that modulates the expression of metformin transporters, were associated with changes in treatment HbA1c (P < 0.01) and metformin secretory clearance (P < 0.05). Population pharmacokinetic modeling further confirmed a 24% reduction in apparent clearance in homozygous carriers of one such variant, rs784888. Genetic variants in other transcription factors, peroxisome proliferator-activated receptor-α and hepatocyte nuclear factor 4-α, were significantly associated with HbA1c change only. Overall, our study highlights the importance of genetic variants in transcription factors as modulators of metformin PK and response.
- Published
- 2014
21. Prenatal caloric restriction enhances DNA methylation and MeCP2 recruitment with reduced murine placental glucose transporter isoform 3 expression
- Author
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Ganguly, Amit, Chen, Yongjun, Shin, Bo-Chul, and Devaskar, Sherin U
- Subjects
Human Genome ,Pediatric Research Initiative ,Preterm ,Low Birth Weight and Health of the Newborn ,Perinatal Period - Conditions Originating in Perinatal Period ,Infant Mortality ,Genetics ,Pediatric ,Aetiology ,2.1 Biological and endogenous factors ,Reproductive health and childbirth ,Animals ,Base Sequence ,Caloric Restriction ,CpG Islands ,DNA Methylation ,DNA Primers ,Female ,Glucose Transporter Type 3 ,Methyltransferases ,Mice ,Placenta ,Pregnancy ,Real-Time Polymerase Chain Reaction ,Fetal growth restriction ,Epigenetics ,Histone deacetylases ,Sp1 transcription factor ,Biochemistry and Cell Biology ,Food Sciences ,Nutrition and Dietetics ,Nutrition & Dietetics - Abstract
Diminished transplacental glucose transport plays an important role in prenatal calorie restriction (CR) induced reduction in fetal growth. Fetal growth restriction (FGR) has an impact in shaping the adult phenotype with transgenerational implications. To understand the mechanisms underlying prenatal CR-induced transplacental glucose transport, we examined the epigenetic regulation of placental glucose transporter (Glut1 and Glut3) expression. We restricted calories by 50% in C57BL6 pregnant mice from gestational days 10 to 19 (CR; n=8) vs. controls (CON; n=8) and observed a 50% diminution in placental Glut3 expression (P
- Published
- 2014
22. DNA methylation status of TBX20 in patients with tetralogy of Fallot
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Juan Gong, Wei Sheng, Duan Ma, Guoying Huang, and Fang Liu
- Subjects
DNA methylation ,TBX20 gene ,Sp1 transcription factor ,Tetralogy of Fallot ,Internal medicine ,RC31-1245 ,Genetics ,QH426-470 - Abstract
Abstract Background TBX20 plays an important role in heart development; however, its epigenetic regulation in the pathogenesis of tetralogy of Fallot (TOF) remains unclear. Methods The methylation levels of the TBX20 promoter region in the right ventricular myocardial tissues of TOF and control samples were measured by the Sequenom MassARRAY platform. Bisulphite-sequencing PCR (BSP) was used to confirm the TBX20 methylation of CpG sites in cells. Dual-luciferase reporter assays were performed to detect the influence of TBX20 methylation and Sp1 transcription factors on gene activity. An electrophoretic mobility shift assay (EMSA) was used to explore the binding of the Sp1 transcription factor to the TBX20 promoter. Results TOF cases had a significantly lower TBX20_M1 methylation level than controls (median methylation: 20.40% vs. 38.73%; p = 0.0047). The Sp1 transcription factor, which binds to Sp1 binding sites in the TBX20_M1 region and promotes TBX20 gene activity, was blocked by the methylation of Sp1 binding sites in normal controls. With decreasing methylation in the TOF cases, the Sp1 transcription factor can bind to its binding site within the TBX20 promoter M1 region and promote TBX20 gene expression. Conclusions Hypomethylation of the TBX20 promoter region was observed in the TOF cases, and the high expression of the TBX20 gene may be caused by activated Sp1 transcription factor binding because of the decreasing methylation at the Sp1 transcription factor binding sites within TBX20_M1.
- Published
- 2019
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23. Genetic selection for context-dependent stochastic phenotypes: Sp1 and TATA mutations increase phenotypic noise in HIV-1 gene expression.
- Author
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Miller-Jensen, Kathryn, Skupsky, Ron, Shah, Priya S, Arkin, Adam P, and Schaffer, David V
- Subjects
Humans ,HIV-1 ,Stochastic Processes ,Transcription ,Genetic ,TATA Box ,Phenotype ,Mutation ,Sp1 Transcription Factor ,Selection ,Genetic ,In Vitro Techniques ,Transcription ,Genetic ,Selection ,Mathematical Sciences ,Biological Sciences ,Information and Computing Sciences ,Bioinformatics - Abstract
The sequence of a promoter within a genome does not uniquely determine gene expression levels and their variability; rather, promoter sequence can additionally interact with its location in the genome, or genomic context, to shape eukaryotic gene expression. Retroviruses, such as human immunodeficiency virus-1 (HIV), integrate their genomes into those of their host and thereby provide a biomedically-relevant model system to quantitatively explore the relationship between promoter sequence, genomic context, and noise-driven variability on viral gene expression. Using an in vitro model of the HIV Tat-mediated positive-feedback loop, we previously demonstrated that fluctuations in viral Tat-transactivating protein levels generate integration-site-dependent, stochastically-driven phenotypes, in which infected cells randomly 'switch' between high and low expressing states in a manner that may be related to viral latency. Here we extended this model and designed a forward genetic screen to systematically identify genetic elements in the HIV LTR promoter that modulate the fraction of genomic integrations that specify 'Switching' phenotypes. Our screen identified mutations in core promoter regions, including Sp1 and TATA transcription factor binding sites, which increased the Switching fraction several fold. By integrating single-cell experiments with computational modeling, we further investigated the mechanism of Switching-fraction enhancement for a selected Sp1 mutation. Our experimental observations demonstrated that the Sp1 mutation both impaired Tat-transactivated expression and also altered basal expression in the absence of Tat. Computational analysis demonstrated that the observed change in basal expression could contribute significantly to the observed increase in viral integrations that specify a Switching phenotype, provided that the selected mutation affected Tat-mediated noise amplification differentially across genomic contexts. Our study thus demonstrates a methodology to identify and characterize promoter elements that affect the distribution of stochastic phenotypes over genomic contexts, and advances our understanding of how promoter mutations may control the frequency of latent HIV infection.
- Published
- 2013
24. Identification of TMZ resistance-associated histone post-translational modifications in glioblastoma using multi-omics data.
- Author
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Ye L, Gu L, Wang Y, Xing H, Li P, Guo X, Wang Y, and Ma W
- Subjects
- Humans, Temozolomide pharmacology, Temozolomide therapeutic use, Histones, Multiomics, Protein Processing, Post-Translational, Sp1 Transcription Factor, Glioblastoma drug therapy, Glioblastoma genetics
- Abstract
Backgroud: Glioblastoma multiforme (GBM) is among the most aggressive cancers, with current treatments limited in efficacy. A significant hurdle in the treatment of GBM is the resistance to the chemotherapeutic agent temozolomide (TMZ). The methylation status of the MGMT promoter has been implicated as a critical biomarker of response to TMZ., Methods: To explore the mechanisms underlying resistance, we developed two TMZ-resistant GBM cell lines through a gradual increase in TMZ exposure. Transcriptome sequencing of TMZ-resistant cell lines revealed that alterations in histone post-translational modifications might be instrumental in conferring TMZ resistance. Subsequently, multi-omics analysis suggests a strong association between histone H3 lysine 9 acetylation (H3K9ac) levels and TMZ resistance., Results: We observed a significant correlation between the expression of H3K9ac and MGMT, particularly in the unmethylated MGMT promoter samples. More importantly, our findings suggest that H3K9ac may enhance MGMT transcription by facilitating the recruitment of the SP1 transcription factor to the MGMT transcription factor binding site. Additionally, by analyzing single-cell transcriptomics data from matched primary and recurrent GBM tumors treated with TMZ, we modeled the molecular shifts occurring upon tumor recurrence. We also noted a reduction in tumor stem cell characteristics, accompanied by an increase in H3K9ac, SP1, and MGMT levels, underscoring the potential role of H3K9ac in tumor relapse following TMZ therapy., Conclusions: The increase in H3K9ac appears to enhance the recruitment of the transcription factor SP1 to its binding sites within the MGMT locus, consequently upregulating MGMT expression and driving TMZ resistance in GBM., (© 2024 The Authors. CNS Neuroscience & Therapeutics published by John Wiley & Sons Ltd.)
- Published
- 2024
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25. SP1-stimulated miR-208a-5p aggravates sepsis-induced myocardial injury via targeting XIAP.
- Author
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Xu LJ, Yang Y, Yuan LF, Liu H, Xu NP, Yang Y, and Huang L
- Subjects
- Animals, Rats, Apoptosis, Lipopolysaccharides pharmacology, Myocytes, Cardiac metabolism, Sp1 Transcription Factor, MicroRNAs metabolism, Sepsis complications, Sepsis genetics, Sepsis metabolism
- Abstract
The development of sepsis can lead to many organ dysfunction and even death. Myocardial injury is one of the serious complications of sepsis leading to death. New evidence suggests that microRNAs (miRNAs) play a critical role in infection myocardial injury. However, the mechanism which miR-208a-5p regulates sepsis-induced myocardial injury remains unclear. To mimic sepsis-induced myocardial injury in vitro, rat primary cardiomyocytes were treated with LPS. Cell viability and apoptosis were tested by CCK-8 and flow cytometry, respectively. The secretion of inflammatory factors was analyzed by ELISA. mRNA and protein levels were detected by RT-qPCR and Western blotting. The interaction among SP1, XIAP and miR-208a-5p was detected using dual luciferase report assay. Ultrasonic analysis and HE staining was performed to observe the effect of miR-208a-5p in sepsis-induced rats. Our findings indicated that miR-208a-5p expression in primary rat cardiomyocytes was increased by LPS. MiR-208a-5p inhibitor reversed LPS-induced cardiomyocytes injury through inhibiting the apoptosis. Furthermore, the inflammatory injury in cardiomyocytes was induced by LPS, which was rescued by miR-208a-5p inhibitor. In addition, downregulation of miR-208a-5p improved LPS-induced sepsis myocardial injury in vivo. Mechanistically, XIAP might be a target gene of miR-208a-5p. SP1 promoted transcription of miR-208a by binding to the miR-208a promoter region. Moreover, silencing of XIAP reversed the regulatory of miR-208a-5p inhibitor on cardiomyocytes injury. To sum up, those findings revealed silencing of miR-208a-5p could alleviate sepsis-induced myocardial injury, which would grant a new process for the treatment of sepsis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)
- Published
- 2024
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26. Regulation of ABO blood group antigen expression by miR‐331‐3p and miR‐1908‐5p during hematopoietic stem cell differentiation.
- Author
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Kronstein‐Wiedemann, Romy, Nowakowska, Paulina, Milanov, Peter, Gubbe, Knut, Seifried, Erhard, Bugert, Peter, Chavakis, Triantafyllos, and Tonn, Torsten
- Subjects
BLOOD transfusion ,ERYTHROCYTES ,GLYCOSYLTRANSFERASES ,HEMATOPOIETIC stem cells ,MICRORNA - Abstract
The ABO blood group system is the most important factor in clinical transfusion medicine and is implicated in a number of human diseases. ABO antigens are not confined to red blood cells (RBCs) and are widely expressed in a variety of human cells and tissues. To date, many alleles with variant ABO expression have been identified and in many cases traced to one of the >250 reported genetic variations in the respective glycosyltransferase. The role of microRNAs (miRNAs) in the regulation of blood group antigens during erythropoiesis has not been addressed, however. Here, we show that miR‐331‐3p and miR‐1908‐5p directly target the mRNA of glycosyltransferases A and B. Expression levels of miR‐331‐3p and miR‐1908‐5p inversely correlated with levels of blood group A antigen. In addition, we found that overexpression of these miRNAs in hematopoietic stem cells led to a significantly reduced number of blood group A antigens per RBC. Simultaneous targeting of the transcription factor (TF) SP1 by miR‐331‐3p further enhanced these effects. The targeting rendered SP1 incapable of binding to the ABO gene promoter, causing further downregulation of blood group A antigen expression by up to 70%. Taken together, expression changes in these miRNAs may account for rare cases of weak A/B phenotypes that genetic variations in the glycosyltransferase coding region cannot explain. These results also suggest an explanation for the disappearance of ABH antigens during carcinogenesis and point to new therapeutic targets in ABO mismatched organ transplantation. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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27. Mithramycin Is a Gene-Selective Sp1 Inhibitor That Identifies a Biological Intersection between Cancer and Neurodegeneration
- Author
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Sleiman, Sama F, Langley, Brett C, Basso, Manuela, Berlin, Jill, Xia, Li, Payappilly, Jimmy B, Kharel, Madan K, Guo, Hengchang, Marsh, J Lawrence, Thompson, Leslie Michels, Mahishi, Lata, Ahuja, Preeti, MacLellan, W Robb, Geschwind, Daniel H, Coppola, Giovanni, Rohr, Jürgen, and Ratan, Rajiv R
- Subjects
Biomedical and Clinical Sciences ,Neurosciences ,Cancer ,Rare Diseases ,Neurodegenerative ,Genetics ,Development of treatments and therapeutic interventions ,5.1 Pharmaceuticals ,2.1 Biological and endogenous factors ,Aetiology ,Neurological ,Analysis of Variance ,Animals ,Animals ,Genetically Modified ,Antibiotics ,Antineoplastic ,Blotting ,Western ,Cell Survival ,Cells ,Cultured ,Cerebral Cortex ,Chromatin Immunoprecipitation ,Drosophila ,Neurons ,Plicamycin ,Rats ,Rats ,Sprague-Dawley ,Sp1 Transcription Factor ,Structure-Activity Relationship ,Medical and Health Sciences ,Psychology and Cognitive Sciences ,Neurology & Neurosurgery - Abstract
Oncogenic transformation of postmitotic neurons triggers cell death, but the identity of genes critical for degeneration remain unclear. The antitumor antibiotic mithramycin prolongs survival of mouse models of Huntington's disease in vivo and inhibits oxidative stress-induced death in cortical neurons in vitro. We had correlated protection by mithramycin with its ability to bind to GC-rich DNA and globally displace Sp1 family transcription factors. To understand how antitumor drugs prevent neurodegeneration, here we use structure-activity relationships of mithramycin analogs to discover that selective DNA-binding inhibition of the drug is necessary for its neuroprotective effect. We identify several genes (Myc, c-Src, Hif1α, and p21(waf1/cip1)) involved in neoplastic transformation, whose altered expression correlates with protective doses of mithramycin or its analogs. Most interestingly, inhibition of one these genes, Myc, is neuroprotective, whereas forced expression of Myc induces Rattus norvegicus neuronal cell death. These results support a model in which cancer cell transformation shares key genetic components with neurodegeneration.
- Published
- 2011
28. cAMP regulates 11β-hydroxysteroid dehydrogenase-2 and Sp1 expression in MLO-Y4/MC3T3-E1 cells.
- Author
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Liu, Di, Wang, Yaoqing, Pan, Zhenyu, Huang, Zhen, and Chen, Fan
- Subjects
- *
TRANSCRIPTION factor Sp1 , *FEMUR head , *ADENYLATE cyclase , *PROTEIN expression , *ENZYME metabolism - Abstract
11β-hydroxysteroid dehydrogenase-2 (11β-HSD2) is one of the key enzymes in glucocorticoid metabolism, which can inactivate local corticosterone and regulate the level of active glucocorticoid in tissues. The expression of 11β-HSD2 and its regulatory pathway serve an important role in the apoptosis of steroid induced osteonecrosis of the femoral head (SANFH). The present study aimed to identify the regulatory effects of cAMP on the expression of Sp1 transcription factor (Sp1) and 11β-HSD2 in osteocytes at the cellular level. Murine long bone osteocyte Y4 (MLO-Y4) clone cells and mouse embryo osteoblast-like (MC3T3-E1) cells were cultured in vitro with adenylate cyclase activator or inhibitor (forskolin and SQ22536, respectively) to investigate the effects of alterations to intracellular cAMP levels. mRNA and protein expression levels of Sp1 and 11β-HSD2 were detected by reverse transcription-quantitative PCR and western blotting, respectively. Compared with the negative control group, the mRNA and protein expression levels of Sp1 were significantly increased in the activation group, whereas Sp1 expression levels were significantly decreased in the inhibition group. Similarly, compared with the negative control group, the mRNA and protein expression levels of 11β-HSD2 were significantly increased in the activator group, but significantly decreased in the inhibitor group. The aforementioned results indicated that intracellular cAMP levels significantly regulated the expression of Sp1 and 11β-HSD2 in mouse osteocytes and osteoblasts. Therefore, the present study suggested a potential therapeutic strategy for the prevention of osteonecrosis of the femoral head. [ABSTRACT FROM AUTHOR]
- Published
- 2020
29. Sequencing of lipoprotein lipase gene in the Mediterranean river buffalo identified novel variants affecting gene expression.
- Author
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Gu, M., Cosenza, G., Gaspa, G., Iannaccone, M., Macciotta, N.P.P., Chemello, G., Di Stasio, L., and Pauciullo, A.
- Subjects
- *
LIPOPROTEIN lipase , *GENE expression , *TRANSCRIPTION factor Sp1 , *GENETIC overexpression , *EXOMES , *WATER buffalo , *SINGLE nucleotide polymorphisms - Abstract
Lipoprotein lipase (LPL) is a key enzyme for lipid metabolism, playing a fundamental role in the composition of fat in adipose tissue and milk. The LPL gene has been seldom investigated in dairy ruminants and barely studied in river buffalo (Bubalus bubalis). The aim of this work was to explore the genetic diversity of LPL and its promoter and to identify functional mutations, using a combined approach based on sequencing, dual-color electrophoretic mobility shift assay, and quantitative PCR. Thirteen consensus sequences for transcription factors were found in the promoter. Eleven SNP were detected, and the attention was focused on the SNP with potential functional effects: g.-446A>G, because the presence of G created a consensus motif for the transcription factor Sp1, and g.107A>G, which was the only exonic SNP. We developed PCR-RFLP methods for genotyping the 2 SNP and calculated the allele frequencies. A strong linkage disequilibrium (D′ = 1; r2 = 0.903) was found between the 2 SNP. The dual-color electrophoretic mobility shift assay demonstrated that only genotype g.-446GG allowed the binding of the Sp1 transcription factor, resulting in overexpression of the gene (~2.5 fold), as confirmed by the quantitative PCR results. Haploinsufficiency is proposed as a regulation mechanism. This study adds further knowledge on the structure of the LPL gene and its expression in river buffalo, with potential effects on milk qualitative and quantitative production. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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30. Sp1-mediated transcription is involved in the induction of leptin by insulin-stimulated glucose metabolism
- Author
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Moreno-Aliaga, MJ, Swarbrick, MM, Lorente-Cebrián, S, Stanhope, KL, Havel, PJ, and Martínez, JA
- Subjects
Obesity ,Nutrition ,Diabetes ,Genetics ,Metabolic and endocrine ,3T3-L1 Cells ,Adipocytes ,Animals ,Cells ,Cultured ,Deoxyglucose ,Gene Expression Regulation ,Glucose ,Insulin ,Leptin ,Male ,Mice ,Okadaic Acid ,Rats ,Rats ,Wistar ,Regulatory Elements ,Transcriptional ,Sp1 Transcription Factor ,Transcription ,Genetic ,Veterinary Sciences ,Clinical Sciences ,Paediatrics and Reproductive Medicine ,Endocrinology & Metabolism - Abstract
We have previously demonstrated that insulin-stimulated glucose metabolism, and not insulin per se, mediates the effects of insulin to increase the transcriptional activity of the leptin promoter in adipocytes. Here, we sought to identify the specific cis-acting DNA elements required for the upregulation of leptin gene transcription in response to insulin-mediated glucose metabolism. To accomplish this, 3T3-L1 cells and primary rat adipocytes were transfected with a series of luciferase reporter genes containing portions of the mouse leptin promoter. Using this method, we identified an element between -135 and -95 bp (relative to the transcriptional start site) that mediated transcription in response to insulin-stimulated glucose metabolism in adipocytes. This effect was abolished by incubation with 2-deoxy-d-glucose, a competitive inhibitor of glucose metabolism. Gel shift electrophoretic mobility shift assays confirmed that the stimulatory effect of insulin-mediated glucose metabolism on leptin transcription was mediated by a previously identified Sp1 site. Consistent with these findings, incubation of primary rat adipocytes with WP631, a specific inhibitor of specificity protein (Sp)1-dependent transcription, inhibited glucose- and insulin-stimulated, but not basal, leptin secretion. Together, these findings support a key role for Sp1 in the transcriptional activation of the leptin gene promoter by insulin-mediated glucose metabolism.
- Published
- 2007
31. E2F7 enhances hepatocellular carcinoma growth by preserving the SP1/SOX4/Anillin axis via repressing miRNA‐383‐5p transcription
- Author
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Fengjie Hao, Nan Wang, Yifan Zhang, Wen Xu, Yongjun Chen, Xiaochun Fei, and Junqing Wang
- Subjects
Cancer Research ,Carcinoma, Hepatocellular ,Sp1 Transcription Factor ,Liver Neoplasms ,Microfilament Proteins ,SOXC Transcription Factors ,Gene Expression Regulation, Neoplastic ,MicroRNAs ,Contractile Proteins ,E2F7 Transcription Factor ,Cell Line, Tumor ,Humans ,Molecular Biology ,Cell Proliferation ,Transcription Factors - Abstract
E2F family participates in most human malignancies by activating the transcription of the cell cycle-related genes. Whereas, as a specifical atypical member of this family, E2F7 was described as a repressor against its downstream genes and exerted oscillatory and controversial functions in cancers. Our previous study identified a molecular interaction promoting hepatocellular carcinoma (HCC) growth induced by SOX4 and Anillin. Meanwhile, we preliminarily identified SP1 as the upstream activator of SOX4. Intriguingly, we observed that the repressive E2F7 presents a remarkable high expression in HCC, and is positively correlated and involved in the same pathway with the potentially SP1/SOX4/Anillin axis. However, their exact interaction or mechanism controlling tumor progress between these genes has not been illustrated. Thus, we focused on this point in this study and attempted to improve the potential regulating axis in HCC cell proliferation and tumor growth for promoting tumor prevention and control. The expression profile of E2F7 in HCC tissues and tumor cells was detected along with the related candidate genes, through real-time quantitative polymerase chain reaction assay, the Western blot analysis, and the immunohistochemistry assay, combined with bioinformatics analysis of the HCC information from the the Cancer Genome Altas and Gene Expression Omnibus data sets. The correlation between E2F7 and HCC patients' clinicopathologic features was explored. Gain-of and loss-of-function assays were conducted both in vitro and in vivo along with the rescue experiment, for revealing the relative genes' functions in HCC progress. The ChIP and the dual-luciferase reporter assays were performed to verify the transcriptional regulating profile between E2F7 and SP1/SOX4/Anillin axis. E2F7 was upregulated in HCC and significantly correlated with SP1/SOX4/Anillin axis. High E2F7 expression is associated with dismal clinicopathologic features and poor survival of the patients. E2F7 depletion potently impaired SP1/SOX4/Anillin expression and significantly inhibited HCC growth. Furthermore, intensive exploration demonstrated that E2F7 preserves high SP1 levels by abrogating miR-383-5p in a transcriptional way. Atypical E2F7 is an important repressive transcription factor commonly upregulated in the HCC environment. E2F7 facilitates HCC growth by repressing miR-383-5p transcription and sequentially promoting SP1/SOX4/Anillin axis. Our findings provide us with probable targets for HCC prevention and therapeutic treatment.
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- 2022
32. Transcription factor Sp1 regulates mitotic chromosome assembly and segregation
- Author
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Samuel Flashner, Michelle Swift, Aislinn Sowash, Alexander N. Fahmy, and Jane Azizkhan-Clifford
- Subjects
Mammals ,Sp1 Transcription Factor ,Chromosome Segregation ,Centromere ,Genetics ,Animals ,Humans ,Mitosis ,Aneuploidy ,Genetics (clinical) ,Transcription Factors - Abstract
Aneuploidy is a pervasive feature of cancer cells that results from chromosome missegregation. Several transcription factors have been associated with aneuploidy; however, no studies to date have demonstrated that mammalian transcription factors directly regulate chromosome segregation during mitosis. Here, we demonstrate that the ubiquitously expressed transcription factor specificity protein 1 (Sp1), which we have previously linked to aneuploidy, has a mitosis-specific role regulating chromosome segregation. We find that Sp1 localizes to mitotic centromeres and auxin-induced rapid Sp1 degradation at mitotic onset results in chromosome segregation errors and aberrant mitotic progression. Furthermore, rapid Sp1 degradation results in anomalous mitotic chromosome assembly characterized by loss of condensin complex I localization to mitotic chromosomes and chromosome condensation defects. Consistent with these defects, Sp1 degradation results in reduced chromosome passenger complex activity and histone H3 serine 10 phosphorylation during mitosis, which is essential for condensin complex I recruitment and chromosome condensation. Together, these data provide the first evidence of a mammalian transcription factor acting specifically during mitosis to regulate chromosome segregation.
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- 2022
33. miR-335-5p regulates the proliferation, migration and phenotypic switching of vascular smooth muscle cells in aortic dissection by directly regulating SP1
- Author
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Runwei, Ma, Dayong, Zhang, Yi, Song, Jichang, Kong, Chunjie, Mu, Pin, Shen, and Wenting, Gui
- Subjects
Sp1 Transcription Factor ,Angiotensin II ,Myocytes, Smooth Muscle ,Becaplermin ,Biophysics ,General Medicine ,Biochemistry ,Muscle, Smooth, Vascular ,Aortic Dissection ,MicroRNAs ,Phenotype ,Cell Movement ,Humans ,Luciferases ,Cells, Cultured ,Cell Proliferation - Abstract
Uncontrolled proliferation, migration and phenotypic switching of vascular smooth muscle cells (VSMCs) are important steps in the development and progression of aortic dissection (AD). The function and potential mechanism of miR-335-5p in the pathogenesis of AD are explored in this study. Specifically, the biological function of miR-335-5p is explored
- Published
- 2022
34. Bone Marrow-Derived Mesenchymal Stem Cell-Derived Exosomes Containing Gli1 Alleviate Microglial Activation and Neuronal Apoptosis In Vitro and in a Mouse Parkinson Disease Model by Direct Inhibition of Sp1 Signaling
- Author
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Yang Cai, Ming-Ming Zhang, Ming Wang, Zhuo-Hang Jiang, and Zhi-Gang Tan
- Subjects
Lipopolysaccharides ,Sp1 Transcription Factor ,Apoptosis ,Mesenchymal Stem Cells ,Parkinson Disease ,General Medicine ,Exosomes ,Zinc Finger Protein GLI1 ,Pathology and Forensic Medicine ,Disease Models, Animal ,Mice ,MicroRNAs ,Neuroblastoma ,Cellular and Molecular Neuroscience ,Neurology ,Bone Marrow ,Animals ,Humans ,Microglia ,Neurology (clinical) - Abstract
This study investigated possible therapeutic effect mechanisms of exosomes from bone marrow-derived mesenchymal stem cells (BMSC) in neuronal and microglial cells and in a Parkinson disease (PD) model. Neuronal SH-SY5Y cells and microglial HMC3 cells were subjected to 1-methyl-4-phenylpyridinium (MPP+) or LPS, respectively. The mRNA and protein expression was assessed using qRT-PCR, Western blotting, and enzyme-linked immunosorbent assay. Cell viability and apoptosis of SH-SY5Y cells were examined using the MTT assay and flow cytometry. Chromatin immunoprecipitation assays were performed to assess the binding relationship between glioma-associated oncogene homolog 1 (Gli1) and the Sp1 transcription factor promoter. BMSC-derived exosomes promoted cell proliferation and inhibited apoptosis in MPP+-treated SH-SY5Y cells and suppressed inflammatory markers in LPS-treated HMC3 cells. Sp1 knockdown decreased SH-SY5Y cell damage and HMC3 immune activation. Gli1 carried by BMSC exosomes directly bound with Sp1 to inhibit Sp1-mediated LRRK2 activation whereas exosomes secreted by Gli1-knockdown in BMSC did not. In a PD mouse model induced with MPTP, BMSC exosomes decreased neuron loss injury and the inflammatory response by inhibiting Sp1 signaling. Thus, BMSC-derived exosomal Gli1 alleviates inflammatory damage and neuronal apoptosis by inhibiting Sp1 in vitro and in vivo. These findings provide the basis for the potential clinical use of BMSC-derived exosomes in PD.
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- 2022
35. Copy number amplification and SP1-activated lncRNA MELTF-AS1 regulates tumorigenesis by driving phase separation of YBX1 to activate ANXA8 in non-small cell lung cancer
- Author
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Xiyi Lu, Jing Wang, Wei Wang, Chenfei Lu, Tianyu Qu, Xuezhi He, Xinyin Liu, Renhua Guo, and Erbao Zhang
- Subjects
Cancer Research ,Lung Neoplasms ,DNA Copy Number Variations ,Carcinogenesis ,Sp1 Transcription Factor ,Gene Expression Regulation, Neoplastic ,Carcinoma, Non-Small-Cell Lung ,Cell Line, Tumor ,Genetics ,Humans ,RNA, Long Noncoding ,Y-Box-Binding Protein 1 ,Molecular Biology ,Cell Proliferation - Abstract
Long non-coding RNAs (lncRNAs) are reported to play key roles in tumorigenesis. However, the mechanisms underlying lncRNA-mediated regulation of RNA-binding protein phase separation in tumorigenesis have not been completely elucidated. In this study, an oncogenic lncRNA MELTF-AS1 was identified using systematic data analysis, screening, and verification. MELTF-AS1 was markedly upregulated in non-small cell lung cancer (NSCLC). High MELTF-AS1 levels were associated with advanced tumor-node-metastasis stage (TNM), high tumor size, and decreased survival time. Functionally, MELTF-AS1 regulated cell proliferation and metastasis in vitro and in vivo. RNA sequencing analysis revealed that MELTF-AS1 knockdown specifically modulated genes associated with cell proliferation, apoptosis, and migration. Mechanistically, at the genome level, copy number amplification promoted MELTF-AS1 expression. At the transcriptional level, the transcription factor SP1 directly activated MELTF-AS1 transcription by binding to its promoter. Furthermore, MELTF-AS1 could directly bind and drive the phase separation of YBX1, which was an RNA-binding protein and involved in tumorigenesis, thus activating ANXA8 transcription and promoting tumorigenesis of NSCLC. Aberrant activation of ANXA8 and promotion of tumorigenesis have been found in a variety of tumors. These novel findings demonstrated the critical role of MELTF-AS1-driven phase separation-mediated transcriptional regulation and provided a potential novel diagnostic and therapeutic target for NSCLC.
- Published
- 2022
36. Sp1 transcription factor represses transcription of phosphatase and tensin homolog to aggravate lung injury in mice with type 2 diabetes mellitus-pulmonary tuberculosis
- Author
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Hongmei, Zhao, Lian, Shi, Xiaohong, Wang, Xiuli, Yu, and Danfeng, Wang
- Subjects
Sp1 Transcription Factor ,PTEN Phosphohydrolase ,Bioengineering ,Lung Injury ,General Medicine ,Applied Microbiology and Biotechnology ,Mice ,Diabetes Mellitus, Type 2 ,Tensins ,Animals ,Humans ,Proto-Oncogene Proteins c-akt ,Tuberculosis, Pulmonary ,Biotechnology - Abstract
Type 2 diabetes mellitus (T2DM) can enhance the risk of mycobacterium tuberculosis (Mtb) infection and aggravate pulmonary tuberculosis (PTB). This study intended to explore the function of phosphatase and tensin homolog (PTEN) in T2DM-PTB and the molecules involved. Mice were treated with streptozotocin to induce T2DM and then infected with Mtb. The mice with T2DM had increased weight, blood glucose level, glucose intolerance and insulin resistance, and increased susceptibility to PTB after Mtb infection. PTEN was significantly downregulated in mice with T2DM-PTB and it had specific predictive value in patients. Overexpression of PTEN improved mouse survival and reduced bacterial load, inflammatory infiltration, cell apoptosis, and fibrosis in lung tissues. Sp1 transcription factor (SP1) was predicted and identified as an upstream regulator of PTEN. SP1 suppressed PTEN transcription. Silencing of SP1 enhanced mouse survival and alleviated the lung injury, and it promoted the M1 polarization of macrophages in murine lung tissues. However, further downregulation of PTEN increased protein kinase B (Akt) phosphorylation and blocked the alleviating roles of SP1 silencing in T2DM-PTB. This study demonstrates that SP1 represses PTEN transcription to promote lung injury in mice with T2DM-PTB through Akt activation.
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- 2022
37. Expression of specificity protein 1 and collagen I in primary pterygial tissues.
- Author
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Bo Jiang, Yong Wang, Zi-Cheng Zhu, and Zhang-You Wu
- Subjects
PROTEIN expression ,COLLAGEN ,TRANSCRIPTION factors ,WESTERN immunoblotting ,TRANSCRIPTION factor Sp1 - Abstract
Copyright of Arquivos Brasileiros de Oftalmologia is the property of Arquivos Brasileiros de Oftalmologia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
- Published
- 2019
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38. The single nucleotide polymorphism g.133A>C in the stearoyl CoA desaturase gene (SCD) promoter affects gene expression and quali-quantitative properties of river buffalo milk.
- Author
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Gu, M., Cosenza, G., Iannaccone, M., Macciotta, N.P.P., Guo, Y., Di Stasio, L., and Pauciullo, A.
- Subjects
- *
DESATURASES , *RUMINANTS , *MILKFAT , *MILK quality , *MILK yield - Abstract
The stearoyl-CoA desaturase (SCD) gene has been investigated in depth in ruminants because of its effect on milk fat composition. In river buffalo, the single nucleotide polymorphism (SNP) g.133A>C in the gene promoter has been associated with milk quality and yield. However, the biological reason for such effects remains unexplored. In this study, we combined mRNA profile analysis, an electromobility shift assay, and quantitative PCR to elucidate the role of this SNP on gene transcription and its effects on milk fat traits. A preliminary genotyping of g.133A>C was carried out on a group of 303 river buffaloes to choose individuals for the downstream applications. Analysis of allele frequencies showed an increase in the minor allele C (0.25) compared with previous findings (0.16). Six animals (2 for each genotype) were chosen for cloning and 216 positive cDNA recombinant clones for SCD (72 per genotype) were analyzed by PCR. All clones showed the same length on agarose gel; therefore, random clones were chosen for sequencing. No qualitative differences were found and all gene transcripts assembled correctly. An electrophoretic mobility shift assay was performed to evaluate the binding of the transcription factor Sp1 to DNA sequences including g.133A>C. Genotype CC showed a higher binding affinity (mean ± standard error of the mean) than genotype AA in 2 different conditions [Enzo buffer (EB), Enzo Life Science Inc., Farmingdale, NY: 201.77 ± 4.06 vs. 141.65 ± 3.77 band intensity values and Poletto buffer (PB): 95.90 ± 1.15 vs. 67.30 ± 2.14 band intensity values]. The subsequent quantitative PCR confirmed the upregulation of the CC genotype compared with the AA and AC genotypes. The association study with milk fat traits revealed a favorable effect of allele C. The heterozygous genotype had the highest values for monounsaturated fatty acids, oleic acid (C18:1 cis-9), polyunsaturated fatty acids, and odd- and branched-chain fatty acids, and the lowest values for saturated fatty acids and atherogenic and thrombogenic indices; the heterozygous genotype differed significantly from the AA genotype. The AC genotype has previously been associated with higher milk yield. Therefore, the g.133A>C SNP is a marker with dual effects and is an interesting candidate for assisted selection programs in river buffalo. These data clarified the biological role of the SNP g.133A>C in the SCD promoter and how it affects gene function, providing important knowledge on the genetic background of lipid metabolism, including the future possibility of selecting alleles with quantitatively or qualitatively favorable effects. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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39. Silencing of specificity protein 1 protects H9c2 cells against lipopolysaccharide-induced injury via binding to the promoter of chemokine CXC receptor 4 and suppressing NF-κB signaling
- Author
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Zhao, Zhu, Guoxiu, Zhang, Dahuan, Li, Xiaojun, Yin, and Tianzhong, Wang
- Subjects
Lipopolysaccharides ,Receptors, CXCR4 ,Sp1 Transcription Factor ,NF-kappa B ,g protein-coupled protein receptor cxc chemokine receptor 4 ,Bioengineering ,General Medicine ,Response Elements ,Applied Microbiology and Biotechnology ,Cell Line ,Rats ,septic myocardial injury ,Animals ,Myocytes, Cardiac ,specificity protein 1 ,nuclear factor kappa b signaling ,TP248.13-248.65 ,Signal Transduction ,Biotechnology - Abstract
G protein-coupled protein receptor CXC chemokine receptor 4 (CXCR4) has been shown to be involved in the development of sepsis; however, it remains unclear whether CXCR4 participates in the septic myocardial injury. In our study, treatment with lipopolysaccharide (LPS) increased the expression of specificity protein 1 (SP1) and CXCR4 in H9c2 cells. Notably, a positive association between SP1 and CXCR4 expression was observed in LPS-treated H9c2 cells, and SP1 positively regulated CXCR4 expression in H9c2 cells. Moreover, silencing of SP1 or CXCR4 suppressed LPS-induced inflammation and cell apoptosis in H9c2 cells, as evidenced by the increase in cell viability and decrease in lactate dehydrogenase release, interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α levels, and caspase-3 activity. Additionally, overexpression of CXCR4 abolished the protective effects of SP1 silencing on LPS-induced injury in H9c2 cells. SP1 was also shown to enhance the promoter activity of CXCR4 by directly binding with the binding motif site - 109/-100 in CXCR4 promoter. Besides, downregulation of SP1 or CXCR4 blocked LPS-induced activation of the NF-кB signaling in H9c2 cells. Furthermore, inhibition of NF-кB signaling by DHMEQ abolished LPS-induced myocardial inflammation and apoptosis. In conclusion, silencing of SP1 protected H9c2 cells against LPS-induced injury by binding to the promoter of CXCR4 and suppressing the NF-κB signaling pathway. Hence, our findings provide evidence that manipulation of SP1 or CXCR4 may be an effective approach to promote prevention or recovery of septic myocardial injury, and thereby, may serve as a potential therapeutic strategy for sepsis.
- Published
- 2022
40. Transcription Factor Specificity Protein 1 Regulates Inflammation and Fibrin Deposition in Nasal Polyps Via the Regulation of microRNA-125b and the Wnt/β-catenin Signaling Pathway
- Author
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Li Song, Xi Wang, Xiangyang Qu, and Chao Lv
- Subjects
Inflammation ,Fibrin ,Sp1 Transcription Factor ,Immunology ,Mice ,MicroRNAs ,Nasal Polyps ,Chronic Disease ,Animals ,Humans ,Immunology and Allergy ,Sinusitis ,Wnt Signaling Pathway ,Rhinitis - Abstract
Nasal polyps (NPs) are multifactorial soft growths inside the nasal passages and are associated with chronic inflammation that originate from the nasal and paranasal sinus mucosae. This study focused on the role of microRNA (miR)-125b and the molecules associated with NP development. Differentially expressed miRNAs between nasal tissues from patients with chronic rhinosinusitis (CRS) with NP (CRSwNP) and CRS without NP (CRSsNP) were screened using microarray analysis. A murine model of CRSwNP was established. The expression of miR-125b in murine tissues was examined using reverse transcription quantitative polymerase chain reaction. Candidate upstream regulators of miR-125b were predicted using bioinformatics tools, and the binding relationship between specificity protein 1 (Sp1) and miR-125b was validated using luciferase and chromatin immunoprecipitation assays. Altered expression of Sp1 and miR-125b was induced to evaluate their relevance to the progression of NPs. miR-125b expression was significantly upregulated in NP tissues from patients with CRSwNP. Sp1 was confirmed as an upstream regulator that promotes miR-125b transcription in NPs. Overexpression of Sp1 reduced levels of d-dimer (an indicator of fibrinogen degradation products) and tissue-type plasminogen activator (t-PA) but increased eosinophil cationic protein and peroxidase levels, as well as the levels of inflammatory factors interleukin-5 (IL-5) and IL-8 in murine NP tissues. However, these trends were reversed after miR-125b downregulation. Sp1 and miR-125b were found to activate the Wnt/β-catenin signaling pathway in NPs. This study demonstrated that Sp1, an upstream transcription factor of miR-125b, accumulates on the miR-125b promoter to activate its transcription, which induces inflammation and fibrin deposition in NP by activating the Wnt/β-catenin axis.
- Published
- 2022
41. The androgen receptor inhibits transcription of GPER1 by preventing Sp1 and Sp3 from binding to the promoters in prostate cancer cells
- Author
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Austin McDermott, KyoungHyun Kim, Susan Kasper, Shuk-Mei Ho, and Yuet-Kin Leung
- Subjects
Male ,GPR30 ,promoter ,Binding Sites ,GPER1 spliced variants ,Sp1 Transcription Factor ,Prostatic Neoplasms ,castration resistant ,Mice ,Sp3 Transcription Factor ,Oncology ,Receptors, Androgen ,Androgens ,Animals ,Humans ,RNA Polymerase II ,RNA, Messenger ,transcription start site ,Research Paper - Abstract
G-1, a GPER1 agonist, was shown to inhibit the growth of castration-resistant mouse xenografts but not their parental androgen-dependent tumors. It is currently unknown how the androgen receptor (AR) represses GPER1 expression. Here, we found that two GPER1 mRNA variants (GPER1v2 and GPER1v4) were transcriptionally repressed, not via transcript destabilization, by the androgen-activated AR. Although no AR binding was found in all active promoters near GPER1, data from promoter assays suggested that both variants' promoters were inhibited by androgen treatment. Site-directed mutagenesis on Sp1/Sp3 binding sites revealed their role in supporting the basal expression of GPER1. Knockdown of Sp1 and Sp3 together but not separately repressed GPER1 expression whereas overexpression of both Sp1 and Sp3 together was required to alleviate AR repression of GPER1. Based on the chromatin immunoprecipitation data, Sp3 was found to bind to the promoters prior to the binding of Sp1 and RNA polymerase II. However, the binding of all three transcription factors was inhibited by DHT treatment. Concordantly, DHT treatment induced nuclear interactions between AR and Sp1 or Sp3. Taken together, these results indicate that AR represses transcription of GPER1 by binding to Sp1 and Sp3 independently to prevent their transactivation of the GPER1 promoters.
- Published
- 2022
42. SP1 Promotes HDAC4 Expression and Inhibits HMGB1 Expression to Reduce Intestinal Barrier Dysfunction, Oxidative Stress, and Inflammatory Response after Sepsis
- Author
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Zhen-Mi Liu, Xi Wang, Chen-Xi Li, Xue-Yan Liu, Xiao-Jing Guo, Yang Li, You-Lian Chen, Hong-Xing Ye, and Huai-Sheng Chen
- Subjects
Lipopolysaccharides ,Mice, Inbred C57BL ,Mice ,Oxidative Stress ,Gastrointestinal Diseases ,Sp1 Transcription Factor ,Sepsis ,Human Umbilical Vein Endothelial Cells ,Animals ,Humans ,Immunology and Allergy ,HMGB1 Protein ,Histone Deacetylases - Abstract
As a serious and elusive syndrome caused by infection, sepsis causes a high rate of mortality around the world. Our investigation aims at exploring the role and possible mechanism of specificity protein-1 (SP1) in the development of sepsis. A mouse model of sepsis was established by cecal ligation perforation, and a cellular model was stimulated by lipopolysaccharide (LPS), followed by determination of the SP1 expression. It was determined that SP1 was poorly expressed in the intestinal tissues of septic mice and LPS-treated cells. Next, we examined the interactions among SP1, histone deacetylase 4 (HDAC4), and high mobility group box 1 (HMGB1) and found that SP1 bound to the HDAC4 promoter to upregulate its expression, thereby promoting the deacetylation of HMGB1. Meanwhile, gain- or loss-of-function approaches were applied to evaluate the intestinal barrier dysfunction, oxidative stress, and inflammatory response. Overexpression of SP1 or underexpression of HMGB1 was observed to reduce intestinal barrier dysfunction, oxidative stress, and inflammatory injury. Collectively, these experimental data provide evidence reporting that SP1 could promote the HDAC4-mediated HMGB1 deacetylation to reduce intestinal barrier dysfunction, oxidative stress, and inflammatory response induced by sepsis, providing a novel therapeutic target for sepsis prevention and treatment.
- Published
- 2022
43. Regulation of lung epithelial cell senescence in smoking-induced COPD/emphysema by microR-125a-5p via Sp1 mediation of SIRT1/HIF-1a
- Author
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Wu, Hao, Ma, Huimin, Wang, Lumin, Zhang, Huazhong, Lu, Lu, Xiao, Tian, Cheng, Cheng, Wang, Peiwen, Yang, Yi, Wu, Meng, Wang, Suhua, Zhang, Jinsong, and Liu, Qizhan
- Subjects
Male ,Sp1 Transcription Factor ,Senescence ,Applied Microbiology and Biotechnology ,Cell Line ,Cigarette Smoking ,Mice ,Pulmonary Disease, Chronic Obstructive ,Sirtuin 1 ,Animals ,Humans ,Lung ,Molecular Biology ,Ecology, Evolution, Behavior and Systematics ,Emphysema ,Mice, Inbred BALB C ,Chronic obstructive pulmonary disease ,Smoking ,Epithelial Cells ,Cell Biology ,respiratory system ,Hypoxia-Inducible Factor 1, alpha Subunit ,respiratory tract diseases ,microRNAs ,Pulmonary Emphysema ,Senescence-Associated Secretory Phenotype ,Research Paper ,Developmental Biology - Abstract
Chronic obstructive pulmonary disease (COPD) affects the health of more than 300 million people worldwide; at present, there is no effective drug to treat COPD. Smoking is the most important risk factor, but the molecular mechanism by which smoking causes the disease is unclear. The senescence of lung epithelial cells is related to development of COPD. Regulation of miRNAs is the main epigenetic mechanism related to aging. β-Galactose staining showed that the lung tissues of smokers have a higher degree of cellular senescence, and the expression of miR-125a-5p is high. This effect is obvious for smokers with COPD/emphysema, and there is a negative correlation between miR-125a-5p levels and values for forced expiratory volume in one second (FEV1)/forced vital capacity (FVC). After Balb/c mice were chronically exposed to various concentrations of cigarette smoke (CS), plethysmography showed that lung function was impaired, lung tissue senescence was increased, and the senescence-associated secretory phenotype (SASP) in bronchoalveolar lavage fluid was increased. For mouse lung epithelial (MLE)-12 cells treated with cigarette smoke extract (CSE), Sp1 and SIRT1 levels were low, HIF-1α acetylation levels were high, and cell senescence and secretion of SASP factors were elevated. Down-regulation of miR-125a-5p or up-regulation of Sp1 reversed these effects. In addition, compared with mice exposed to CS, knockdown of miR-125a-5p reduced lung epithelial cell senescence and COPD/emphysema. Therefore, in smoking-induced COPD, elevated miR-125a-5p participates in the senescence of lung epithelial cells through Sp1/SIRT1/HIF-1α. These findings provide evidence related to the pathogenesis of COPD/emphysema caused by chronic smoking.
- Published
- 2022
44. Regulatory Effect of Methylation of the Porcine AQP3 Gene Promoter Region on Its Expression Level and Porcine Epidemic Diarrhea Virus Resistance
- Author
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Jia-Yun Wu, Fang Wang, Zheng-Chang Wu, Sheng-Long Wu, and Wen-Bin Bao
- Subjects
pig ,PEDV ,AQP3 gene ,methylation ,Sp1 transcription factor ,Genetics ,QH426-470 - Abstract
As an important carrier for intestinal secretion and water absorption, aquaporin 3 (AQP3) is closely related to diarrhea. In this study, we investigated the mechanisms of AQP3 gene expression regulation in porcine epidemic diarrhea virus (PEDV)-induced diarrhea confirmed by PCR amplification and sequencing. Evaluation of intestinal pathology showed that diarrhea caused by PEDV infection destroyed the intestinal barrier of piglets. qPCR analysis showed that AQP3 expression in the small intestine of PEDV-infected piglets was extremely significantly decreased. qPCR and Bisulfite sequencing PCR revealed an increase in the methylation levels of both CpG islands in the AQP3 promoter region in the jejunum of PEDV-infected piglets. The methylation of mC-20 and mC-10 sites within the two CpG islands showed a significant negative correlation with AQP3 expression. Chromatin Co-Immunoprecipitation (ChIP)-PCR showed that the Sp1 transcription factor was bound to the AQP3 promoter region containing these two CpG sites. AQP3 expression was also extremely significantly reduced in Sp1-inhibited IPEC-J2 cells, indicating that abnormal methylation at the mC-20 site of CpG1 and the mC-10 site of CpG2 reduces its expression in PEDV-infected piglet jejunum by inhibiting the binding of Sp1 to the AQP3 promoter. These findings provide a theoretical basis for further functional studies of porcine AQP3.
- Published
- 2020
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45. Knockdown of versican 1 in lung fibroblasts aggravates Lipopolysaccharide-induced acute inflammation through up-regulation of the SP1-Toll-like Receptor 2-NF-κB Axis: a potential barrier to promising Versican-targeted therapy.
- Author
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Xu, Lulu, Hu, Weiping, Zhang, Jing, and Qu, Jieming
- Subjects
- *
FIBROBLASTS , *CELL migration , *LIPOPOLYSACCHARIDES , *CELL communication , *CYTOKINE release syndrome , *TOLL-like receptors , *ANGIOTENSIN II - Abstract
• knockdown of Versican 1 could bring dramatically increased expression of inflammatory factors. • LPS-induced inflammation is induced by knockdown of V1 via the SP1-TLR2-NF-κB axis. • Increased expression of V1 might be protective in acute inflammation. Versican participates in various pathological processes like inflammation and fibrosis and is a potential therapeutic target for inflammatory diseases. Versican 1 (V1) has increased expression in inflammatory diseases, but its role is unclear. We explored the effects of V1 on acute lung inflammation to determine whether targeting V1 had therapeutic potential. Human fetal lung fibroblast (HFL1) was transfected with or without V1-inhibiting lentivirus and treated with LPS. The expression levels of inflammatory cytokines, V1, cellular signaling pathway and Toll-like receptors (TLRs) were detected by qPCR, ELISA and western blot. The migration and adhesion of neutrophils and monocytes to HFL1s were performed. The activity of transcriptional factors was determined by dual-luciferase reporter assay. Inflammatory factors increased dramatically and continuously with V1 knockdown and LPS stimulation (P < 0.01), orchestrating migration of inflammatory cells and an enhanced inflammatory reaction. V1-knockdown increased TLR2 (P < 0.01) and activated the NF-κB pathway, which was partially reversed with a TLR2 neutralizing antibody and an NF-κB inhibitor. Explosion of LPS-induced inflammation was induced by knockdown of V1 via the SP1-TLR2-NF-κB axis. Increased expression of V1 might be protective in acute inflammation, and an infection-induced cytokine storm might be a severe complication of V1-targeted interventions. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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46. miR-590-3 and SP1 Promote Neuronal Apoptosis in Patients with Alzheimer’s Disease via AMPK Signaling Pathway
- Author
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Yanqun Cao, Xiangxiang Tan, Quzhe Lu, Kai Huang, Xiaoer Tang, and Zhiming He
- Subjects
MicroRNAs ,Article Subject ,Alzheimer Disease ,Sp1 Transcription Factor ,Medical technology ,Humans ,Apoptosis ,Radiology, Nuclear Medicine and imaging ,AMP-Activated Protein Kinases ,R855-855.5 ,Research Article ,Signal Transduction - Abstract
Alzheimer’s disease (AD) is a progressive neurological degenerative illness with a hidden onset. Its pathogenesis is complicated, although with molecular biology research on cancer and targeted research on pathogenic mechanisms, good progress has not yet been made. Therefore, this work built a multifactor-driven neuronal apoptosis dysfunction module for the purpose of probing its underlying pathogenic mechanisms. We performed differential expression analysis, coexpression analysis, enrichment analysis, and hypergeometric tests to calculate the underlying regulatory effects of multifactors on the modules by the way of the whole gene expression profile of AD and identify a series of ncRNA (miR-320a) and TF (NFKB1). Additionally, we screened 10 modules corresponding to the Hub gene, which tend to regulate the physiological progress of inflammation, regulation of autophagy, cerebral cortex neuron differentiation, glial cell apoptotic, and so on. Meanwhile, Alzheimer’s disease is triggered by signaling pathways such as the MPK signaling pathway. In this study, a dysfunction module is utilized to verify that miR-590-3 and SP1 motility factors can regulate neurons in Alzheimer’s disease through the MPK signaling pathway, not only providing new insights into the pathogenesis of Alzheimer’s disease but also laying a solid theoretical foundation for the biologists to further cure Alzheimer’s disease.
- Published
- 2021
47. Specificity protein 1/microRNA-92b forms a feedback loop promoting the migration and invasion of head and neck squamous cell carcinoma
- Author
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Pai Pang, Hui Fang, Hong Wu, Song Wang, Minda Liu, Shan Jin, Zhongzheng Qi, Zhenning Li, Fayu Liu, and Changfu Sun
- Subjects
Male ,Sp1 Transcription Factor ,Mice, Nude ,Bioengineering ,head and neck squamous cell carcinoma ,Applied Microbiology and Biotechnology ,Cell Movement ,Cell Line, Tumor ,Biomarkers, Tumor ,Animals ,Humans ,Neoplasm Invasiveness ,Neoplasm Metastasis ,specificity protein 1 ,Cell Proliferation ,Feedback, Physiological ,Squamous Cell Carcinoma of Head and Neck ,General Medicine ,Middle Aged ,microrna-92b ,Prognosis ,migration and invasion ,Gene Expression Regulation, Neoplastic ,MicroRNAs ,stomatognathic diseases ,Head and Neck Neoplasms ,Multivariate Analysis ,Disease Progression ,Female ,feedback loop ,TP248.13-248.65 ,Research Article ,Research Paper ,Biotechnology - Abstract
In this study we report a novel specificity protein 1 (SP1)/microRNA-92b (miR-92b) feedback loop regulating the migration and invasion of head and neck squamous cell carcinoma (HNSCC). Microarray and real-time Polymerase Chain Reaction (PCR) were used to detect gene expression in HNSCC tissues and cell lines. Transwell migration, invasion, wound healing and cell counting kit – 8 (CCK-8) cell assays were used to compare cell migration, invasion and proliferation abilities. Chromatin Immunoprecipitation (ChIP) assays were used to detect SP1 binding to the miR-92b promoter. Western blot was used to detect protein levels. An in vivo tumorigenesis experiment was used to evaluate the effect of SP1 knockdown on tumor growth and protein levels were evaluated by immunohistochemistry. We found that the miR-92b expression level was elevated in HNSCC primary focus tissue compared with adjacent normal tissue, and a higher level of miR-92b was related to a higher clinical stage and worse prognosis of HNSCC patients. MiR-92b and SP1 mutually promoted each expression and cooperatively facilitated the migration, invasion and proliferation of HNSCC cells. A decreased level of SP1/miR-92b resulted in a restraint of in vivo tumor growth. In conclusion, our results suggest that the SP1/miR-92b feedback loop generally promotes HNSCC invasion and metastasis, thus presenting a possible therapeutic target in the treatment of HNSCC patients.
- Published
- 2021
48. Long Noncoding RNA NEAT1 Knockdown Ameliorates 1-Methyl-4-Phenylpyridine–Induced Cell Injury Through MicroRNA-519a-3p/SP1 Axis in Parkinson Disease
- Author
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Shuihua Wang, Bohai Xiong, Zhang Li, Qinli Wen, Yu Xiaoli, and Ouyang Xiaochun
- Subjects
1-Methyl-4-phenylpyridinium ,Gene knockdown ,Dose-Response Relationship, Drug ,Cell Survival ,Herbicides ,Sp1 Transcription Factor ,business.industry ,Parkinson Disease ,Paraspeckle ,Cell biology ,MicroRNAs ,Downregulation and upregulation ,Apoptosis ,Cell Line, Tumor ,Gene Knockdown Techniques ,microRNA ,Humans ,Gene silencing ,Medicine ,RNA, Long Noncoding ,Surgery ,Neurology (clinical) ,Viability assay ,business ,Transcription factor - Abstract
Background Parkinson disease is a neurodegenerative disease and is characterized by resting tremor, dementia, and gait disorder. Previous studies have indicated that long noncoding RNA participates in the regulation of the pathogenesis of Parkinson disease. The study aimed to reveal the effects of long noncoding RNA nuclear paraspeckle assembly transcript 1 (NEAT1) on 1-methyl-4-phenylpyridine (MPP+)–induced human neuroblastoma cell injury and the underlying mechanism. Methods The expressions of NEAT1, microRNA (miR)-519a-3p, and transcription factor specific protein 1 (SP1) were detected by quantitative real-time polymerase chain reaction. The protein expressions of SP1 and inflammation-related factors were determined by Western blot. Cell viability was determined by 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell apoptosis was investigated by flow cytometry analysis. The targeting relationship between miR-519a-3p and NEAT1 or SP1 was predicted by starBase online database and verified by a dual-luciferase reporter assay. Results NEAT1 and SP1 expressions were significantly upregulated, whereas miR-519a-3p was downregulated in MPP+-treated neuroblastoma cells in a dose- and time-dependent manner when compared with control groups. NEAT1 knockdown restrained MPP+-induced repression of cell viability and promotion of cell apoptosis and inflammation. Additionally, NEAT1 served as a sponge of miR-519a-3p and regulated MPP+-caused cell injury by interacting with miR-519a-3p. Also, SP1, a target gene of miR-519a-3p, rescued miR-519a-3p–mediated actions under MPP+ treatment. Importantly, NEAT1 stimulated SP1 expression through interaction with miR-519a-3p. Conclusions NEAT1 silencing protected against MPP+-induced neuroblastoma cell injury by regulating the miR-519a-3p/SP1 pathway. This finding provides a novel direction for the development of therapeutic strategies for Parkinson disease.
- Published
- 2021
49. Enhanced Sp1/YY1 Expression Directs CBS Transcription to Mediate VEGF-Stimulated Pregnancy-Dependent H 2 S Production in Human Uterine Artery Endothelial Cells
- Author
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Dong-bao Chen and Jin Bai
- Subjects
Adult ,Vascular Endothelial Growth Factor A ,Transcription, Genetic ,Sp1 Transcription Factor ,Article ,chemistry.chemical_compound ,Pregnancy ,Transcription (biology) ,medicine.artery ,Internal Medicine ,medicine ,Humans ,Hydrogen Sulfide ,Promoter Regions, Genetic ,Uterine artery ,YY1 Transcription Factor ,biology ,YY1 ,Endothelial Cells ,Middle Aged ,medicine.disease ,Cystathionine beta synthase ,Cell biology ,Vascular endothelial growth factor ,Endothelial stem cell ,Uterine Artery ,Vascular endothelial growth factor A ,chemistry ,embryonic structures ,biology.protein ,Female - Abstract
Pregnancy and VEGF (vascular endothelial growth factor) stimulate uterine artery endothelial cell (UAEC) hydrogen sulfide production via selectively upregulating CBS (cystathionine β-synthase) but not CSE (cystathionine γ-lyase) expression. This study was conducted to determine the mechanisms by which VEGF utilizes to stimulate pregnancy-dependent upregulation of CBS and hydrogen sulfide production in human UAEC. The proximal human CBS promoter contains 4 Sp1 (specificity protein 1; a/b/c/d) sites and 1 YY1 (Yin Yang 1) site; luciferase assays using reporter genes driven by human CBS promoter with a series of 5′-deletions identified a promoter sequence (−574 to −394) containing Sp1d and the YY1 sites critical for basal and VEGF-stimulated CBS promoter activation. VEGF stimulated pregnancy-dependent recruitment of Sp1 to Sp1d and YY1 to YY1 and also recruited YY1 to Sp1c and increased Sp1/YY1 association in pregnant human UAEC, suggesting formation of a Sp1/YY1 complex at the Sp1c site. Endothelial Sp1 and YY1 proteins were significantly greater in pregnant than nonpregnant human uterine artery. VEGF stimulated pregnancy-dependent Sp1 and YY1 protein expression in vitro. Treatment with Sp1 and YY1 siRNAs completely blocked Sp1/YY1-mediated pregnancy-dependent CBS protein upregulation and hydrogen sulfide production by VEGF in human UAEC. VEGF did not trans -activate CSE promoter or increase CSE expression, and Sp1/YY1 knockdown did not affect CSE expression in human UAEC. Thus, pregnancy augments EC Sp1 and YY1 expression and promotes the recruitment of Sp1/YY1 to their DNA-binding sequences in proximal human CBS promoter to upregulate CBS transcription, underlying a novel mechanism to mediate VEGF-stimulated pregnancy-dependent endothelial hydrogen sulfide production in the human uterine artery.
- Published
- 2021
50. The Acetylation Modification of SP1 Regulates the Protein Stability in Silkworm
- Author
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Zhengying You, Dan Wang, Zihan Sun, Caiying Jiang, Jiao Lv, Qing Sheng, Zuoming Nie, Yue Liu, and Yafei Ma
- Subjects
Sp1 transcription factor ,biology ,Protein Stability ,Chemistry ,Immunoprecipitation ,fungi ,Acetylation ,Bioengineering ,General Medicine ,Bombyx ,Applied Microbiology and Biotechnology ,Biochemistry ,Blot ,Ubiquitin ,Downregulation and upregulation ,Tandem Mass Spectrometry ,Proteome ,Gene expression ,biology.protein ,Animals ,Protein Processing, Post-Translational ,Molecular Biology ,Biotechnology - Abstract
Acetylation is a highly conservative and reversible post-translational modification. Acetylation modification can regulate gene expression by altering protein function and is widely identified in an increasing number of species. Previously, the acetylated proteome of silkworm was identified by combining acetylated polypeptide enrichment with nano-HPLC/MS/MS; the identification revealed that the SP proteins (SPs) were high acetylated. In this study, the acetylation of SP1, one of the SPs, was further confirmed using immunoprecipitation (IP) and Western blotting. Then, we found the acetylation could upregulate SP1 protein expression by enhancing the protein stability. Further research found that the acetylation of SP1 protein can competitively inhibit its ubiquitination and thus improve the stability and cell accumulation of SP1 protein by inhibiting the ubiquitin-mediated proteasome degradation pathway. This result provides a basis for acetylation to regulate the nutrient storage and utilization of silkworm.
- Published
- 2021
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