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Oral submucous fibrosis is a progressive oral mucosal condition that is characterized by inflammation and persistent fibrosis. Epithelial Mesenchymal Transition is a crucial molecular event that contributes to tumor progression and fibrosis, with ZEB 1 and its effect on E-cadherin expression being key molecules in the process. Thera are no tissue level studies of these molecules in oral submucous fibrosis. To evaluate the immunohistochemical expression of epithelial mesenchymal transition markers E-cadherin and ZEB1 in oral submucous fibrosis. A total of 30 cases of Oral submucous fibrosis (15 Early OSMF and 15 Advanced OSMF) classified based on the histopathological features were included in the study. Immunohistochemistry was done using two markers i.e. E-cadherin and ZEB1. The difference in the expression of E-Cadherin and ZEB1 among histo-pathological grades of OSMF was done by Mann-Whitney U test. A slight reduction in the E-cadherin expression was noted in Oral submucous fibrosis but marked enhanced expression of ZEB1 was seen in the connective tissue of OSMF. An increase in intensity and percentage of positivity of ZEB 1 expression in connective tissue was observed in advanced cases as compared to early OSMF. This can be attributed to role of ZEB1 in mediating EMT via transdifferentiation of fibroblast into myofibroblast and thus predispose to fibrosis in OSMF. [Display omitted] [ABSTRACT FROM AUTHOR]

Mutations in the splicing factor 3b subunit 1 (SF3B1) gene are frequent in myelodysplastic neoplasms (MDS). Because the splicing process is involved in the production of circular RNAs (circRNAs), we investigated the impact of SF3B1 mutations on circRNA processing. Using RNA sequencing, we measured circRNA expression in CD34+ bone marrow MDS cells. We defined circRNAs deregulated in a heterogeneous group of MDS patients and described increased circRNA formation in higher‐risk MDS. We showed that the presence of SF3B1 mutations did not affect the global production of circRNAs; however, deregulation of specific circRNAs was observed. Particularly, we demonstrated that strong upregulation of circRNAs processed from the zinc finger E‐box binding homeobox 1 (ZEB1) transcription factor; this upregulation was exclusive to SF3B1‐mutated patients and was not observed in those with mutations in other splicing factors or other recurrently mutated genes, or with other clinical variables. Furthermore, we focused on the most upregulated ZEB1‐circRNA, hsa_circ_0000228, and, by its knockdown, we demonstrated that its expression is related to mitochondrial activity. Using microRNA analyses, we proposed miR‐1248 as a direct target of hsa_circ_0000228. To conclude, we demonstrated that mutated SF3B1 leads to deregulation of ZEB1‐circRNAs, potentially contributing to the defects in mitochondrial metabolism observed in SF3B1‐mutated MDS. [ABSTRACT FROM AUTHOR]

Cervical cancer (CC) is the most common type of gynecological malignancy in women, and targeting stem cells and inhibiting tumor stem cell-like properties of CC remains an important field of research as an attempt to improve treatment outcomes. This study focused on Ubiquitin-specific-processing Protease 43 (USP43), a member of the deubiquitinase (DUBs) family known to play a role in tumor progression, and analysis of the The Cancer Genome Atlas (TCGA) data and survival computations revealed that USP43 was highly expressed in CC and correlated with poor prognosis. However, the role of USP43 in CC has been under-reported, and its underlying mechanism remains unclear. To investigate the effect and mechanism of USP43, its expression in CC cells and tissues were examined, and the results showed that it was significantly upregulated. Subsequently, knockdown experiments revealed that reducing USP43 expression suppressed CC cell proliferation, and depleting USP43 inhibited the stem cell-like properties of CC cells and impaired their migration abilities. Further investigations indicated that USP43 promoted Zinc finger E-box binding protein 1 (ZEB1)-induced activation of Extracellular regulatory kinase 1/2 (ERK1/2) signaling in CC. Based on these findings, we propose that USP43 could serve as a promising target for CC. [ABSTRACT FROM AUTHOR]

The epithelial-mesenchymal transition (EMT) is a crucial morphological event that occurs during epithelial tumor progression. Snail and ZEB1/2 (ZEB1 and ZEB2), known as EMT transcription factors, are key regulators of this transition. ZEB1/2 are positively correlated with EMT phenotypes and the aggressiveness of cancers. On the contrary, Snail is also correlated with the aggressiveness of cancers, but is not correlated with the expression of EMT marker proteins. Snail is induced by transforming growth factor-b (TGF-b), a well-known inducer of EMT, in various cancer cells. Interestingly, Snail induction by TGF-b is markedly enhanced by active Ras signals. Thus, cancer cells harboring an active Ras mutation exhibit a drastic induction of Snail by TGF-b alone. Here, we found that members of the E26 transformation-specific (Ets) transcription factor family, Ets1 and Ets2, contribute to the upregulation of both Snail and ZEB1/2. Snail induction by TGF-b and active Ras is dramatically inhibited using siRNAs against both Ets1 and Ets2 together, but not on their own; in addition, siRNAs against both Ets1 and Ets2 also downregulate the constitutive expression of Snail and ZEB1/2 in cancer cells. Examination of several alternatively spliced variants of Ets1 revealed that p54-Ets1, which includes exon VII, but not p42-Ets1, which excludes exon VII, regulates the expression of the EMT transcription factors, suggesting that Ets1 is a crucial molecule for regulating Snail and ZEB1/2, and thus cancer progression is mediated through post-translational modification of the exon VII domain. [ABSTRACT FROM AUTHOR]

Aims: We focused on the detailed functions of circ-ABCB10 in cervical cancer (CC) development and its mechanisms. Background: The increasing findings have proposed the central roles of circular RNAs (circRNAs) in the tumorigenesis of various human cancers. Circ-ABCB10 displays promising oncogenic effect in several tumors. Methods: Circ-ABCB10 and miR-128-3p production levels in CC tissues and cells were tested through RT-qPCR. The association of circ-ABCB10 expression with clinicopathologic parameters of CC patients was statistically analyzed. Cell proliferation, invasion, apoptosis, and epithelial-mesenchymal transition (EMT) were evaluated by MTT, transwell invasion assays, flow cytometry analyses, and western blot examination of EMT markers. The binding activity between miR-128-3p and circ-ABCB10 or zinc finger E-box binding homeobox 1 (ZEB1) was explored through pull-down assay or luciferase reporter assay. The influence of circ-ABCB10 on CC tumorigenesis was evaluated by in vivo xenograft experiments. Results: The elevated circ-ABCB10 expression was determined in CC tissues and cells. Moreover, higher production level of circ-ABCB10 was close related to lymph-node metastasis, Federation of Gynecology and Obstetrics (FIGO) stage, and tumor size in CC patients. Loss of circ-ABCB10 weakened cell proliferative and invasive abilities, inhibited EMT, and induced apoptosis in CC. Loss of circ-ABCB10 inhibited ZEB1 expression by serving as a sponge of miR-128-3p in CC cells. Circ-ABCB10 sponged miR-128-3p to enhance cell proliferation, invasion, EMT and inhibit apoptosis in CC cells. Xenograft tumor assays confirmed that circ-ABCB10 knockdown inhibited CC tumor growth. Conclusion: Our study suggests that circ-ABCB10 depletion inhibits proliferation, invasion and EMT and promotes apoptosis of cervical cancer cells through miR-128-3p/ZEB1 axis and represses CC tumor growth. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Osteosarcoma (OS) is an adolescent idiopathic malignancy with a poor prognosis. Accumulating evidence has verified that long non-coding RNAs (lncRNAs) were implicated in the initiation and development of various tumors. We aimed to clarify the functions and underlying mechanism of lncRNA PCAT-1 in OS progression. PATIENTS AND METHODS: RT-qPCR was performed to examine the relative expressions of PCAT-1, miR-508-3p and ZEB1 in OS tissues or cells. The proliferation capacities of OS cells with different transfection were detected by CCK-8 assays. Transwell assays were carried out to determine the functions of PCAT-1 and miR-508-3p in OS cell migration and invasion. Moreover, bioinformatical analysis and Luciferase reporter assay were applied to verify the association between PCAT-1 and miR-508-3p, miR-508-3p and ZEB1. RESULTS: Data of current study revealed that PCAT-1 was markedly upregulated in OS, which indicated poor prognosis of OS patients. CCK-8 and transwell assays indicated that PCAT-1 upregulation could promote OS cell proliferation, invasion and migration. Additionally, we found that miR-508-3p was a direct target of PCAT-1, and PCAT-1 regulated the development of OS via decreasing miR-508-3p and activating its target gene ZEB1. CONCLUSIONS: All data demonstrated that PCAT-1 promoted OS progression, and miR-508-3p/ZEB1 axis was implicated in the functional roles of PCAT-1 in OS, suggesting that PCAT-1/miR-508-3p/ZEB1 might serve as candidate therapeutic targets for OS patients. [ABSTRACT FROM AUTHOR]

Preeclampsia (PE) is characterized by the impaired invasive ability of trophocytes, which can be modulated by microRNAs (miRs). In the current study, the effects of rhynchophylline (Rhy) on the viability and invasive ability of trophocytes were explored by focusing on miR-141-3p/ZEB1 axis. The level of miR-141-3p was modulated in human trophocytes and the changes in cell viability, apoptosis, invasive ability, and ZEB1 level were detected. Then the trophocytes with miR-141-3p overexpression were treated with Rhy and the effects on trophocyte phenotypes were assessed. The induced miR-141-3p level suppressed cell viability, induced apoptosis, and inhibited invasion and ZEB1 level in trophocytes. The treatment of Rhy restored the viability and invasive ability of trophocytes under the overexpression of miR-141-3p, indicating the protective effects of Rhy on trophocytes. The findings in the current study highlighted the protective effects of Rhy on trophocytes during PE progression, which was associated with the inhibition of miR-141-3p. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The purpose of this study was to uncover the potential role of non-coding RNA activated by DNA damage (lncRNA NORAD) in the disease progression of diabetic cardiomyopathy (DCM) and the underlying mechanism. MATERIALS AND METHODS: Cell viability, 5-Ethynyl-2’-deoxyuridine (EdU)-positive ratio and apoptotic rate in human cardiomyocyte cell line AC16 undergoing treatment of normal-level (NG) or high-level glucose (HG) were assessed at first. NORAD level in HG-induced AC16 cells at different time points was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Subsequently, cell viability, EdU-positive ratio, and apoptotic rate in HG-induced AC16 cells overexpressing NORAD were evaluated. Next, subcellular distribution of NORAD was examined, and Dual-Luciferase reporter gene assay was performed to clarify the interaction among NORAD, miRNA-150-5p, and ZEB1. At last, rescue experiments were conducted to clarify the role of NORAD/miRNA-150-5p/ZEB1 axis in influencing the proliferation and apoptosis in HG-induced AC16 cells. RESULTS: Results revealed that HG treatment suppressed the proliferative ability and stimulated apoptosis in AC16 cells. Besides, NORAD was time-dependently downregulated in HG-induced AC16 cells, and it was mainly distributed in cytoplasm. In addition, the overexpression of NORAD enhanced proliferative ability, attenuated apoptosis, and increased Bcl2/Bax ratio in HG-induced AC16 cells. Finally, NORAD/miRNA-150-5p/ZEB1 axis was verified to protect the malignant progression of DCM. CONCLUSIONS: NORAD is upregulated under high-level glucose treatment. Overexpression of NORAD protects DCM development via miRNA150-5p/ZEB1 axis. [ABSTRACT FROM AUTHOR]

Long noncoding RNA (lncRNA) have emerged as crucial regulators for a myriad of biological processes, and perturbations in their cellular expression levels have often been associated with cancer pathogenesis. In this study, we identified AATBC (apoptosis‐associated transcript in bladder cancer, LOC284837) as a novel lncRNA. AATBC was found to be highly expressed in nasopharyngeal carcinoma (NPC), and increased AATBC expression was associated with poor survival in patients with NPC. Furthermore, AATBC promoted migration and invasion of NPC cells in vitro, as well as metastasis in vivo. AATBC upregulated the expression of the desmosome‐associated protein pinin (PNN) through miR‐1237‐3p sponging. In turn, PNN interacted with the epithelial–mesenchymal transition (EMT) activator ZEB1 and upregulated ZEB1 expression to promote EMT in NPC cells. Collectively, our results indicate that AATBC promotes NPC progression through the miR‐1237‐3p–PNN–ZEB1 axis. Our findings indicate AATBC as a potential prognostic biomarker or therapeutic target in NPC. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma (HCC) is one of the most lethal cancers owing to the high metastasis rate. The molecular mechanism underlying HCC progression remains unclear. We aimed to explore the function and mechanism of action of insulin-like growth factor binding protein 2 (IGFBP2) in HCC. Expression of IGFBP2 was evaluated with western blotting and reverse transcription polymerase chain reaction (RT-PCR). Loss- and gain-function assays were conducted to evaluate the effects of IGFBP2 on HCC cell proliferation, migration, and invasion. Signaling pathways were screened with a dual-fluorescein reporting system, and levels of epithelial and mesenchymal markers were measured after altering IGFBP2 expression. Cell fractionation analysis was conducted to evaluate the nuclear translocation of p65. IGFBP2 expression was upregulated in HCC tissues, predicted worse prognosis, and was associated with strong metastatic potentials. IGFBP2 depletion significantly inhibited HCC cell proliferation, migration, and invasion, whereas IGFBP2 overexpression showed reverse phenotypes. The underlying mechanism involved IGFBP2-mediated nuclear localization of p65, which activated nuclear factor kappa B (NF-κB) and zinc finger E-Box binding homeobox 1 (ZEB1) transcription via binding to the gene promoter. This study for the first time identifies IGFBP2 as a novel therapeutic target in HCC that activates the NF-κB-ZEB1 signaling axis and promotes HCC tumorigenesis. [ABSTRACT FROM AUTHOR]

Although uncommon, pulmonary sarcomatoid carcinoma carries a worse prognosis due to poor chemotherapeutic response. Currently, a histologic spindle and/or giant cell component indicates sarcomatoid differentiation, with zinc E-box binding homeobox 1 (ZEB1) implicated in promoting epithelial-mesenchymal transition. However, diagnostic use of ZEB1 in limited specimens, including cell block (CB) preparations, remains unclear. Pulmonary sarcomatoid (SARC, n = 15), typical (TC, n = 10) and atypical carcinoid (AC, n = 10), small cell (SCLC, n = 8) and large cell neuroendocrine carcinoma (LCNEC, n = 9), squamous cell carcinoma (SQ, n = 7), and adenocarcinoma (ADC, n = 7) CBs along with 69 SARCs, 20 TCs, 21 ACs, 9 SCLCs, 10 LCNECs, 71 SQs, 402 ADCs, 16 large cell carcinoma (LCC) and 17 other thoracic tumor (OT) surgical specimens between 2007 and 2018 were retrieved. ZEB1 (Sigma Aldrich, St. Louis, Mo and Novus Biological, Centennial, Colo) immunohistochemistry was graded 1+ to 3+, with ≥1+ and >5% staining considered positive. Nuclear ZEB1 was seen in 80% SARC (12/15), 0% TC (0/10), 0% AC (0/10), 12.5% SCLC (1/8) and 11.1% LCNEC (1/9), 0% SQ (0/7), and 0% ADC (0/7) CBs. In surgical specimens, 75.4% SARCs (52/69), 0% TCs (0/20), 0% ACs (0/21), 11.1% SCLCs (1/9), 30% LCNECs (3/10), 0% SQs (0/71), 0.2% ADCs (1/402), 12.5% LCCs (2/16), and 11.8% OTs (2/17) demonstrated ZEB1. ZEB1 sensitivity and specificity in cytology and surgical specimens were 80% and 96.1%, and 75.4% and 98.1%, respectively. ZEB1 is sensitive and highly specific for pulmonary sarcomatoid carcinoma in limited cytologic and surgical specimens. Diagnostic pitfalls include high-grade neuroendocrine tumors and large cell carcinoma, which are resolvable by morphologic considerations. [ABSTRACT FROM AUTHOR]

OBJECTIVE: This research aims to investigate the level of microRNA-1236-3p in breast cancer (BCa) tissues and to further investigate its possible mechanism in the progression of BCa. PATIENTS AND METHODS: The level of microRNA-1236-3p in BCa tissues and adjacent tissues was detected by quantitative Real-time polymerase chain reaction (qRT-PCR). Regulatory effects of microRNA-1236-3p on cell proliferation and invasion were detected by cell counting kit-8 (CCK-8) and transwell assay. The binding relationship between microRNA-1236-3p and zinc-finger E-box binding homeobox (ZEB1) was examined by the dual-luciferase reporter gene assay. Finally, rescue experiments were conducted to verify the potential role of microRNA-1236-3p/ ZEB1 axis in BCa. RESULTS: MicroRNA-1236-3p was downregulated in BCa tissues relative to adjacent tissues, and the similar trend was shown in BCa cell lines. Overexpression of microRNA-1236-3p in MDA-MB-231 and MCF-7 cells inhibited proliferation and attenuated invasiveness, while knockdown of microRNA-1236-3p had an opposite effect. Dual-luciferase reporter gene assay and qRT-PCR results showed that microRNA-1236-3p could target ZEB1 to degrade it. Overexpression of ZEB1 in BCa cells can partially reverse the effect of overexpressed miR-1236-3p on cell proliferative and invasive abilities. CONCLUSIONS: MicroRNA-1236-3p could inhibit the growth and metastasis of BCa cells by inhibiting ZEB1 expression, suggesting that microRNA-1236-3p may be a potential therapeutic target for BCa. [ABSTRACT FROM AUTHOR]

OBJECTIVE: In order to investigate the role of the zinc finger E-box-binding homeobox 1 (ZEB1) expression in the incidence of non-small cell lung cancer (NSCLC), the effects of the ZEB1 expression on the pathogenesis and prognosis of NSCLC were investigated. PATIENTS AND METHODS: Correlations of the expression of ZEB1 in 88 clinical patients with NSCLC with clinicopathological features were determined. The patients were followed up for 60 months to study the correlation between the ZEB1 expression and the prognosis of NSCLC patients. To further explore the role of the ZEB1 expression in the incidence of NSCLC, the expressions of ZEB1 and E-cadherin in three NSCLC cell lines (A549, NCI-H1299 and NCI-H1975) and human normal lung epithelial BESA-2B cell line were measured, and the cell invasion ability was detected. The role of ZEB1 in NSCLC cell invasion was verified through the knockdown of ZEB1 by the short hairpin ribonucleic acids (shRNAs). In addition, its effects on the proliferation and apoptosis of NSCLC cells were confirmed by the overexpression of ZEB1. RESULTS: The expression of ZEB1 in NSCLC tissues was remarkably higher than that in normal tissues, and the expression level of ZEB1 was significantly related to the cancer stage and tumor size. The lower the expression of ZEB1 was, the higher the overall survival rate and the longer the survival time would be. The expression of ZEB1 was negatively correlated with that of E-cadherin in cell lines, and ZEB1-shRNAs markedly reduced the invasion ability of NSCLC cells. The overexpression of ZEB1 resulted in an increase in the proliferation activity and a significant decrease in the apoptosis of A549 cells. CONCLUSIONS: The expression of ZEB1 is closely related to the incidence and prognosis of NSCLC. Increased expression of ZEB1 is helpful for promoting the invasion of NSCLC and enhancing the proliferation activity of NSCLC. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To explore the role of hsa-microRNA-99b/let-7e/microRNA-125a cluster in the progression of liver cancer and its possible regulatory mechanisms. PATIENTS AND METHODS: Ten liver cancer tissues were randomly selected and matched with normal liver tissue samples. The mRNA expression levels of miR-99b, let-7e and miR-125a were detected by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). The three miRNA mimics were transfected alone or together into hepatoma cells SMMC-7721; at the same time, the knockdown of the three miRNAs was also performed. Then the cell invasive and migratory abilities were examined through transwell assay. Bioinformatics analysis was used to detect the potential transcription factors that bind to the hsa-microRNA-99b/let-7e/microRNA-125a promoter sequence, and the binding of the two was verified by the Luciferase reporting assay. The level of hsa-microRNA-99b/let-7e/microRNA-125a mature mRNA was detected after ZEB1 was inhibited by ZEB1 siRNA. Meanwhile, after interfering with Drosha and ZEB1, the expression level of hsa-microRNA-99b/let-7e/microRNA-125a of the primary transcript was examined. Rescue experiments were carried out to assess the role of hsa-microRNA-99b/let-7e/microRNA-125a in the ZEB1 regulation of invasive cell capacity. RESULTS: The mRNA levels of microRNA-99b, let-7e, and microRNA-125a in 10 selected hepatocarcinoma tissues were significantly higher than those in the matched paracancerous tissues. After overexpressing the three miRNA mimics either alone or together, cell invasive and migratory abilities were extensively enhanced, and vice versa. It was found that there is a binding site in the upstream sequence of the promoter region of the hsa-microRNA-99b/let- 7e/microRNA-125a cluster for ZEB1. The Luciferase reporter gene results showed an increase in the Luciferase activity of the cells transfected with E-box element wild-type sequence, while mutant E-box element group did not change. After knocking out ZEB1, the levels of mature microRNA-99b, let-7e and microRNA-125a were reduced. However, when DROSHA was knocked out, the levels of immature microRNA-99b, let- 7e and microRNA-125a were increased, while simultaneous knocking out ZEB1 reversed this effect. Besides, the invasive ability of SMMC-7721 cells decreased after KEB1 was knocked down, while the opposite result was observed after transfection of hsa-microRNA-99b/let-7e/microRNA-125a alone or together. CONCLUSIONS: Hsa-microRNA-99b/let-7e/microRNA- 125a cluster is highly-expressed in hepatocarcinoma, and its expression can be regulated by ZEB1. In addition, the overexpression of this cluster can promote the invasion of liver cancer cells and advance liver cancer progression. [ABSTRACT FROM AUTHOR]

Objective: Uterine carcinosarcoma (UCS) is a highly aggressive neoplasm that is composed of an intricate admixture of carcinomatous and sarcomatous elements. The relationship between UCS and the epithelial to mesenchymal transition (EMT) has been reported. In this study, we examined how expression of E-cadherin was associated with the expression of EMT-related proteins in UCS. Methods: UCS samples were histologically divided into three components: carcinomatous, transitional, and sarcomatous regions. Next, we examined the expression of E-cadherin and EMT-related proteins, including SNAI2, ZEB1, and TWIST1, in each component of the UCS using immunohistochemistry. The expression score was determined by combining the staining intensity and staining area of the target cells. Results: The expression score of E-cadherin was significantly lower in transitional and sarcomatous components than in the carcinomatous component. In addition, a significant difference in the low expression score of E-cadherin between transitional and sarcomatous components (transitional > sarcomatous components) was found. There were significant differences between the expression scores of ZEB1 in the three components (sarcomatous > transitional > carcinomatous components). However, no difference in the expression of TWIST1 between the components was found. Conversely, the expression level of SNAI2 was higher in sarcomatous or transitional components than in the carcinomatous component. However, a significant difference between the transitional and sarcomatous components was not detected. Conclusion: These results suggest that the EMT plays an essential role in the pathogenesis of UCS. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Recently, studies have identified that microRNAs (miRNAs) are novel regulators for gene expression in tumor progression including breast cancer. The aim of the study is to investigate the clinical significance and underlying functions between miR-508-3p expression and triple-negative breast cancer (TNBC) development. PATIENTS AND METHODS: Quantitative Real-time PCR (QRT-PCR) was performed to determine the expression of miR-508-3p in 54 pairs of TNBC specimens and adjacent non-tumor tissues. The association between miR-508-3p expression and clinicopathological factors was assessed using x²-test. Transwell invasion assays were used to assess cell invasion ability. Luciferase reporter assay, Western blot analyses and qRT-PCR were performed to demonstrate ZEB1 was a direct target of miR-508-3p. RESULTS: We demonstrated that miR-508-3p expression was remarkably decreased in TNBC tissues and cells. Lower miR-508-3p expression significantly associated with lymph node metastasis and distant metastasis in TNBC patients (p < 0.05). Ectopic expression of miR-508-3p significantly suppressed cell invasion ability of TNBC. MiR-508-3p overexpression suppressed cell epithelial-mesenchymal transition (EMT) phenomenon of TNBC by upregulating E-cadherin expression, but downregulating Vimentin expression. In addition, we revealed that ZEB1 was a direct target of miR-508-3p in TNBC cells. MiR-508-3p significantly suppressed cell EMT process by regulating ZEB1 expression. CONCLUSIONS: W e f ound t hat m iR-508-3p may be a potential therapeutic target of TNBC. [ABSTRACT FROM AUTHOR]

Background/purpose: MicroRNA-200c (miR-200c) recently emerged as an important regulator of tumorigenesis and cancer metastasis, however, its role in regulating oral submucous fibrosis (OSF) remains unknown. In this study, we investigated the functional role of miR-200c in myofibroblastic differentiation activity and identified its potential target.Methods: qRT-PCR was applied to assess the expression of miR-200c in OSF tissues and fibrotic buccal mucosal fibroblasts (fBMFs). Arecoline, a major areca nut alkaloid, was utilized to explore whether the expression of miR-200c would alter following stimulation. Collagen gel contraction, migration and invasion capabilities were examined in arecoline-stimulated BMFs as wells as in fBMFs. Luciferase reporter assay was conducted to show the relationship between miR-200c and ZEB1.Results: Our results showed that the expression of miR-200c was downregulated in OSF specimen and fBMFs. Arecoline treatment dose-dependently reduced the relative expression of miR-200c in normal BMFs. Overexpression of miR-200c impeded the arecoline-induced collagen gel contraction, migration, invasion and wound healing capacities. Moreover, ectopic expression of miR-200c in fBMFs successfully reduced the increased collagen gel contractility and invasion abilities. Our results demonstrated that ZEB1 was a direct target of miR-200c, and overexpression of miR-200c inhibited the expression of ZEB1 and α-SMA.Conclusion: These findings suggest that downregulation of miR-200c in OSF may be involved in the pathogenesis of areca nut-associated OSF through regulation of ZEB1. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To investigate the effect of BCL6 on methotrexate-resistant children with acute B lymphoblastic leukemia (B-ALL) and its underlying mechanism. PATIENTS AND METHODS: Bone marrow samples of B-ALL children diagnosed at The Children & Women's Healthcare of Laiwu City from June 2014 to February 2017 were collected. Subjects were assigned into methotrexate-resistant group (n=8) and non-resistant group (n=32) according to the follow-up outcome. Quantitative Real-Time Polymerase Chain Reaction (qRTPCR) was performed to detect expressions of BCL6 and E-cadherin in bone marrow tissues. Cell cycle and apoptosis of methotrexate-resistant B-ALL cells (BALL-1/MTXR cells) were detected after overexpression or inhibition of BCL6, respectively. Western blot was performed to determine the protein levels of E-cadherin and BCL6. The direct binding of BCL6 in the ZEB1 promoter region was verified by the ChIP (chromatin immunoprecipitation) assay. RESULTS: QRT-PCR showed a higher BCL6 expression in bone marrow samples of methotrexate-resistant group than that of the non-resistant group. Moreover, BCL6 was upregulated in BALL-1/MTXR cells than that of untreated B-ALL cells. After knockdown of BCL6 expression, we observed a decreased IC50, increased apoptosis, and arrested the cell cycle in BALL-1/ MTXR cells. In addition, increased expression of E-cadherin was found in BALL-1/MTXR cells, which could be reversed by ZEB1 overexpression. ChIP assay suggested that BCL6 bound to the promoter region of ZEB1, so as to promote ZEB1 expression. CONCLUSIONS: BCL6 is overexpressed in the bone marrow of methotrexate-resistant children with B-ALL, which is capable of attenuating the sensitivity of B-ALL to methotrexate via promoting ZEB1 expression. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To examine the potential mechanisms implicating miR-200c and epithelial-mesenchymal transition (EMT) in oral squamous carcinoma (OSC). MATERIALS AND METHODS: 32 pairs of OSC tissue samples and matched para-carcinoma normal tissue from patients undergoing routine surgery in the Xuzhou Stomatological Hospital from 2014-2016. HOC313 cells were cultured and transfected with miR-200c mimics and scrambled mimics. Cell migration, invasion assays, Luciferase reporter assay, and Western blot assay were conducted. RESULTS: miR-200c was downregulated in OSC tissues compared with adjacent normal tissues (n=32). miR-200c knockdown in the human oral cancer cell line HOC313 significantly suppressed cell invasion and migration, indicating the ability to inhibit tumor progression. Luciferase reporter assay indicated that miR-200c directly bound to the 3'-untranslated regions (3ʹ-UTR) of Zinc finger E-box-binding homeobox (ZEB1) directly. Moreover, miR-200c significantly inhibited HOC313 cell EMT via negatively regulating ZEB1 protein expression. CONCLUSIONS: MiR-200c plays a pivotal role in controlling OSC metastasis via inhibiting EMT, which provides potential therapeutic targets for OSC. [ABSTRACT FROM AUTHOR]

Long non‐coding RNA (lncRNA) is responsible for a diverse range of cellular functions, such as transcriptional and translational regulation and variance in gene expression. The lncRNA CASC15 (cancer susceptibility candidate 15) is a long intergenic non‐coding RNA (lincRNA) locus in chromosome 6p22.3. Previous research shows that lncRNA CASC15 is implicated in the biological behaviors of several cancers such as neuroblastoma and melanoma. Here, we aimed to explore in detail how CASC15 contributes to the growth of gastric cancer (GC). As predicted, the expression of CASC15 was enriched in GC tissues and cell lines as compared with healthy tissues and cells using qRT‐PCR. The Kaplan–Meier method was used to demonstrate that high expression of CASC15 is linked to a poor prognosis for patients suffering from GC. Additionally, functional experiments proved that the down‐ or up‐regulation of CASC15 inhibited or facilitated cell proliferation via the induction of cell cycle arrest and apoptosis, and also suppressed or accelerated cell migration and invasion by affecting the progression of the epithelial‐to‐mesenchymal transition (EMT). In vivo experiments showed that the knockdown of CASC15 lessened the tumor volume and weight and influenced the EMT process. This was confirmed by western blot assays and immunohistochemistry, indicating impaired metastatic ability in nude mice. CASC15 involvement in the tumorigenesis of GC occurs when CASC15 interacts with EZH2 and WDR5 to modulate CDKN1A in nucleus. Additionally, the knockdown of CASC15 triggered the silencing of ZEB1 in cytoplasm, which was shown to be associated with the competitive binding of CASC15 to miR‐33a‐5p. [ABSTRACT FROM AUTHOR]

Epithelial-to-mesenchymal transition (EMT) has an active role in the malignant progression of epithelial tumor cells. The aim of the study was to identify the existence of the EMT mechanism in brain metastasis. Tumors from 29 patients with brain metastases were assessed in terms of the immunoexpression of EMT-related factors including Slug, ZEB1, ZEB2, and E-cadherin in tumor cells and the surrounding mesenchymal stromal cells. The results were compared between primary tumors and their matched metastatic brain tumors. Analysis of tumor cell expression showed that Slug, ZEB1, or ZEB2 expression was found in more than 10% of the neoplastic cells in the metastatic lesions of 17 cases (59%) and in the primary lesions of 7 cases (24%, P = 0.02). The expression level of ZEB2 was negatively correlated with that of E-cadherin ( P = 0.05). There were no differences in the tumoral expression levels of Slug, ZEB1, or ZEB2 among the primary organs. Analysis of stromal cell expression revealed a global increase in ZEB1 and ZEB2 expression levels with metastases ( P < 0.0001). Quantitative analysis confirmed that messenger RNA expression of ZEB1 and ZEB2 was elevated in metastatic lesions. The increased expression of EMT-related factors in brain metastasis was found not only in tumor cells, but also in tumor-associated stromal cells. Our results suggest that EMT-related factors play a role as a facilitator of brain metastasis. [ABSTRACT FROM AUTHOR]

OBJECTIVE: This research focuses on the influence of linc-ITGB1 on the metastasis of hepatocellular carcinoma and further explores its underlying mechanism. PATIENTS AND METHODS: A total of 70 hepatocellular carcinoma patients were chosen for our study. RT-qPCR was used for detecting the expression level of linc-ITGB1 in their cancer tissues. Moreover, the expression level of linc-ITGB1 was also detected in hepatocellular carcinoma cell lines. Furthermore, whether linc-ITGB1 could affect the migrated and invaded ability of hepatocellular carcinoma cells was determined by wound healing assay and transwell assay. We further explored the potential mechanism by RT-qPCR and Western blot assay. RESULTS: L inc-ITGB1 expression level in hepatocellular carcinoma tissues was remarkably higher than that in adjacent tissues. Moreover, migrated and invaded ability of hepatocellular carcinoma cells was inhibited through knockdown of linc-ITGB1. Further study revealed that silenced linc-ITGB1 inhibited the expression of ZEB1 and then suppressed epithelial to mesenchymal transition (EMT), which was important during the metastasis of hepatocellular carcinoma. Moreover, the inhibition of cell invasion by silenced linc-ITGB1 could be rescued through overexpression of ZEB1 in hepatocellular carcinoma. CONCLUSIONS: The results indicate that linc-ITGB1, a novel oncogene in tumorigenesis, could promote the metastasis and EMT via ZEB1, which may offer a possible therapeutic target in hepatocellular carcinoma. [ABSTRACT FROM AUTHOR]

Melanoma progression involves multiple molecular pathways. In this study, we explored the effect of immunoglobulin heavy constant gamma 1 (IGHG1) on melanoma progression. IGHG1, zinc finger E-box binding homeobox 1 (ZEB1), and cleavage stimulation factor subunit 2 tau variant (CSTF2T) expression levels were measured by quantitative reverse-transcription polymerase chain reaction and western blot. BALB/c nude mice were subcutaneously injected with A375 cells to develop a transplantation tumor model, 4 weeks after which the tumor tissues were collected. Cell proliferation, cell invasion, and cell migration of A375 and SK-MEL-28 cells were measured after gain- and loss-of-function of IGHG1, ZEB1, and CSTF2T. Binding of ZEB1 to the IGHG1 promoter was assayed by chromatin immunoprecipitation and dual-luciferase reporter gene assays, while binding of CSTF2T to ZEB1 mRNA was investigated by RNA immunoprecipitation and RNA pull-down assays. IGHG1, ZEB1, and CSTF2T were all highly expressed in human melanoma cell lines. In A375 and SK-MEL-28 cells, ZEB1 binds directly to the IGHG1 promoter. ZEB1 silencing reduced IGHG1 expression and melanoma cell proliferative, invasive, and migratory properties, and these cellular effects were nullified by overexpression of IGHG1. CSTF2T binds directly to ZEB1 mRNA. Silencing of CSTF2T diminished ZEB1 expression and restrained melanoma cell proliferative, invasive, and migratory capabilities. The impacts of CSTF2T silencing on melanoma cells were counteracted by concomitant overexpression of ZEB1. CSTF2T knockdown reduced tumor volume and weight in nude mice. The CSTF2T–ZEB1–IGHG1 axis promotes melanoma cell proliferation and invasion. • IGHG1 is highly expressed in melanoma cells; ZEB1 binds to the promoter of IGHG1. • ZEB1 upregulates IGHG1 to boost melanoma cell proliferation, migration, and invasion. • CSTF2T binds to ZEB1 to promote melanoma cell proliferation, migration, and invasion. • CSTF2T promotes the in the tumor growth of nude mice. • CSTF2T-ZEB1-IGHG1 modulates melanoma cell proliferation, migration, and invasion. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Prostate cancer is a kind of malignancy with high occurrence in the male urogenital system. However, the mechanism of the occurrence, the progression, and the metastasis of prostate cancer are still unclear. Searching for the effective molecule target is of great significance to improve the curative effect on prostate cancer. Zinc finger E box binding protein-1 (ZEB1) protein is a member of the zinc finger transcription factor family that participates in the embryonic development and formation. ZEB1 was found to be involved in the occurrence and in the development of multiple cancers, while its role in prostate cancer still needs elucidation. MATERIALS AND METHODS: Normal prostate cell line PC-3M and prostate cancer cell line DU145 were cultured in vitro and transfected by ZEB1 siRNA. ZEB1 mRNA and protein expressions were detected by real-time PCR and Western blot assay. Cell proliferation was determined by using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Cell migration was evaluated by transwell assay. Cell apoptosis was evaluated by caspase-3 activity. The impact of ZEB1 on extracellular signal-regulated kinase 1 and 2 (ERK1/2) signaling pathway was assessed by Western blot assay. RESULTS: ZEB1 expression significantly increased in DU145 cells compared with PC-3M cells (p<0.05). ZEB1 mRNA and protein obviously declined, cell proliferation inhibited, cell invasion suppressed, and Caspase-3 activity enhanced in DU145 cells after ZEB1 siRNA transfection (p<0.05). ZEB1 siRNA markedly decreased ERK1/2 phosphorylation in DU145 cells compared with control (p<0.05). CONCLUSIONS: Inhibition of ZEB1 promoted prostate cancer apoptosis, restrained proliferation, and suppressed invasion through down-regulating ERK1/2 signaling pathway. [ABSTRACT FROM AUTHOR]

Zinc finger E-box binding protein 1 ( ZEB1) and ZEB2 induce epithelial-mesenchymal transition ( EMT) and enhance cancer progression. However, the global view of transcriptional regulation by ZEB1 and ZEB2 is yet to be elucidated. Here, we identified a ZEB1-regulated inflammatory phenotype in breast cancer cells using chromatin immunoprecipitation sequencing and RNA sequencing, followed by gene set enrichment analysis ( GSEA) of ZEB1-bound genes. Knockdown of ZEB1 and/or ZEB2 resulted in the downregulation of genes encoding inflammatory cytokines related to poor prognosis in patients with cancer, including IL6 and IL8, therefore suggesting that ZEB1 and ZEB2 have similar functions in terms of the regulation of production of inflammatory cytokines. Antibody array and ELISA experiments confirmed that ZEB1 controlled the production of the IL-6 and IL-8 proteins. The secretory proteins regulated by ZEB1 enhanced breast cancer cell proliferation and tumor growth. ZEB1 expression in breast cancer cells also affected the growth of fibroblasts in cell culture, and the accumulation of myeloid-derived suppressor cells in tumors in vivo. These findings provide insight into the role of ZEB1 in the progression of cancer, mediated by inflammatory cytokines, along with the initiation of EMT. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To investigate changes in the peripheral blood mRNA levels of oxytocin receptor (OXTR) and Zinc finger E-box-binding homeobox 1 (ZEB1) before and after progesterone dosing in pregnant women with threatened premature labor. PATIENTS AND METHODS: Blood samples were collected from 30 healthy pregnant women with 28- to 33+6-week gestational age and singleton pregnancy (group A) and from 30 pregnant women with singleton pregnancy and threatened premature labor before and 48 hours after progesterone dosing (groups B and C, respectively) for quantitative real-time reverse transcription polymerase chain reaction (qRTPCR) assay to assess the OXTR and ZEB1 mRNA levels. RESULTS: The OXTR mRNA level was higher in the group B than in the groups A and C and the ZEB1 mRNA level was lower in the group B than in the groups A and C. Notably, no significant difference was found in the mRNA level of OXTR or ZEB1 between group A and group C. CONCLUSIONS: The peripheral blood mRNA level of OXTR was increased and that of ZEB1 was decreased in pregnant women with threatened premature labor. Progesterone helped to maintain pregnancy by readjusting the mRNA levels of OXTR and ZEB1. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Osteosarcoma is one common malignant bone tumors, as it frequently has invasion, metastasis and recurrence, causing unfavorable prognosis of patients. Osteosarcoma has complicated pathogenesis, which has not been elucidated fully. Therefore, the identification of effective molecular target of osteosarcoma onset can help to improve treatment efficacy and prognosis of osteosarcoma. Zinc finger E-box binding homeobox 1 (ZEB1) protein is one member of zinc finger E-box binding protein family, and participates in embryonic genesis and development. A recent study found the participation of ZEB1 in mediating multiple tumor onset and its up-regulation of osteosarcoma. The regulatory mechanism of ZEB1 in osteosarcoma has not been illustrated yet. MATERIALS AND METHODS: In vitro cultured osteosarcoma MG-63 cells were transfected with ZEB1 siRNA. Real-time PCR and Western blot were tested for ZEB1 mRNA/protein expression. MTT was used to test MG-63 cell proliferation, whilst cell invasion was used to describe the effect of ZEB1 on MG-63 cells. Caspase-3 activity assay was employed to test MG-63 cell apoptosis. Western blot was employed to detect nuclear factor kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) protein expression. RESULTS: After transfecting with ZEB1 siRNA, MG-63 cell proliferation or invasion was inhibited accompanied with lower ZEB1 mRNA/protein expression. Caspase3 activity was also increased after transfection (p < 0.05), along with down-regulation of NF-κB and iNOS proteins in MG-63 cells (p < 0.05). CONCLUSIONS: Inhibition of ZEB1 can facilitate osteosarcoma cell apoptosis and inhibit cell proliferation or invasion via down-regulating NF- κB/iNOS signal pathway [ABSTRACT FROM AUTHOR]

Docetaxel is the main treatment for advanced castration-resistant prostate cancer; however, resistance eventually occurs. The development of intratumoral drug-resistant subpopulations possessing a cancer stem cell ( CSC) morphology is an emerging mechanism of docetaxel resistance, a process driven by epithelial-mesenchymal transition ( EMT). This study characterised EMT in docetaxel-resistant sublines through increased invasion, MMP-1 production and ZEB1 and ZEB2 expression. We also present evidence for differential EMT across PC-3 and DU145 in vitro resistance models as characterised by differential migration, cell colony scattering and susceptibility to the CSC inhibitor salinomycin. si RNA manipulation of ZEB1 and ZEB2 in PC-3 and DU145 docetaxel-resistant sublines identified ZEB1, through its transcriptional repression of E-cadherin, to be a driver of both EMT and docetaxel resistance. The clinical relevance of ZEB1 was also determined through immunohistochemical tissue microarray assessment, revealing significantly increased ZEB1 expression in prostate tumours following docetaxel treatment. This study presents evidence for a role of ZEB1, through its transcriptional repression of E-cadherin to be a driver of both EMT and docetaxel resistance in docetaxel-resistant prostate cancer. In addition, this study highlights the heterogeneity of prostate cancer and in turn emphasises the complexity of the clinical management of docetaxel-resistant prostate cancer. [ABSTRACT FROM AUTHOR]

Objective Oral squamous cell carcinoma is characterized by diverse histopathological features, tumorigenesis, and invasion. Gingiva squamous cell carcinoma (GSCC) is characterized by easily invading the jawbone. Epithelial–mesenchymal transition (EMT) is a crucial event for invasion of cancer cells and progression of carcinogenesis, inducing stem-like properties in epithelial cells. Here, expression of BMI1, which controls stem cell self-renewal, and ZEB1, a transcription factor that regulates EMT, was evaluated for its role in EMT and invasion of GSCC. Materials and methods We analyzed 36 gingival specimens (24 carcinomas and 12 dysplasias) by measuring protein and mRNA expression levels of BMI1, ZEB1, vimentin, and E-cadherin, and examining associations between their expression and clinicopathological parameters. Results Compared with adjacent normal squamous epithelium, E-cadherin expression was decreased significantly ( P < 0.05) and vimentin expression levels were increased significantly ( P < 0.05) at the invasive front, which are indicative of EMT in GSCC. Vimentin expression was significantly increased in poorly differentiated carcinoma ( P = 0.027), and vimentin and BMI1 expression was significantly increased in grade 4 mode of invasion ( P < 0.05). Furthermore, elevated levels of BMI1 were accompanied by downregulation of E-cadherin. A significant correlation was found between E-cadherin and BMI1 expression levels ( P = 0.014) and between BMI1 and ZEB1 mRNA expression ( P = 0.034). Conclusion The present study shows that the expression levels of several biomarkers of gingiva carcinogenesis have unique characteristics that differ from those in tongue carcinogenesis. BMI1 and ZEB1 may be important factors associated with EMT in the invasion and progression of GSCC. [ABSTRACT FROM AUTHOR]

OBJECTIVE: This study aimed to investigate the expression of miR-200c and miR-141 in glioma tissues and cell lines and then to study their regulative effect on ZEB1 expression and on glioma cell growth and migration. MATERIALS AND METHODS: QRT-PCR analysis was performed to detect miR-200c and miR-141 expression in 10 paired glioma tissues and adjacent normal tissues from patients with glioblastoma multiforme (GBM) and in glioma cell lines. U87 and U251 cells were transfected with miR-200c mimics, miR-141 mimics or ZEB1 siRNA respectively. ZEB1 expression was detected qRT-PCR andWestern blot assay. MTT assay, flow cytometry and wound healing assay were performed to examine the tumor suppressive effects of the miR-200c/miR-141-ZEB1 axis on glioma cells. RESULTS: Both miR-200c and miR-141 were significantly lower in glioma tissues than in adjacent normal tissues. The glioma cell lines, including U87, U251 and A172 also had significantly decreased miR-200c and miR-141 expression than normal tissues. ZEB1 expression had at least two-fold increase in glioma tissues than in normal tissues. Both miR-200c and miR-141 could significantly induce ZEB1 mRNA degradation and suppress ZEB1 protein expression. ZEB1 siRNA presented similar growth and migration inhibiting and apoptosis inducing effect to miR-200c and miR-141 mimics in U87 cells. CONCLUSIONS: MiR-200c and miR-141 are significantly downregulated in glioma tissues and cell lines and can significantly induce ZEB1 mRNA degradation and suppress ZEB1 protein expression in the cells. ZEB1 is a functional downstream target of miR-200c and miR-141 in inhibiting glioma cell growth and migration. [ABSTRACT FROM AUTHOR]

Abstract A 60-year-old man was admitted to our hospital with non-small cell lung cancer (NSCLC). Imaging and pathological studies revealed NSCLC, not otherwise specified (NOS), at clinical stage T3N1M0 stage IIIA. We started radiotherapy alone because of obstructive pneumonia and end-stage renal disease, but the tumors progressed rapidly and resulted in death due to air obstruction by pharyngeal metastasis. The cancer was diagnosed as pleomorphic carcinoma in an autopsy. Viable lung tumor cells, which were resistant to radiotherapy, and the pharyngeal metastasis had mesenchymal phenotypes and expressed ZEB1 but not SNAI1. These observations indicated that ZEB1-associated epithelial-mesenchymal transition has malignant features including resistance to radiotherapy and aggressive metastatic potential. ZEB1-associated EMT may be an important mechanism to understand the pathophysiology of pleomorphic carcinoma. [ABSTRACT FROM AUTHOR]

Zinc finger E-box binding homeobox factor 1 (ZEB1) is a transcription factor involved in the epithelial to mesenchymal transition (EMT) process of metaplastic breast cancer (MBC). This study aimed to assess the expression of ZEB1 in MBC and explore its association with clinicopathological factors and prognosis. We analyzed the immunohistochemical expression of ZEB1 in 50 MBC tissue samples. ZEB1 was overexpressed in 36% (18/50) of cases. ZEB1 overexpression was significantly correlated to fibromatosis-like and spindle cell sarcoma subtypes (P < 0.001) and tended to be correlated to metastatic status (P = 0.069). Using the Kaplan-Meier method, ZEB1 expression was significantly associated with poor 5-years overall survival (OS) (P = 0.001) and relapse-free survival (RFS) (P = 0.0001). The multivariate Cox regression analysis showed that ZEB1 positive remained a significantly independent adverse prognostic factor for RFS and OS (HR = 4.9 [2.14–11.53]; P < 0.0001) and (HR = 4 [1.05–15.18]; P = 0.042), while Vimentin was an independent poor prognostic factor only for RFS (HR = 5.69 [1.79–18.11], P = 0.003). Our results indicated that ZEB1 and Vimentin overexpression might serve as adverse prognostic factors and potential therapeutic targets for MBC patients. • Overexpression of ZEB1 protein was detected in 36% of metaplastic breast cancer (MBC). • ZEB1 overexpression was significantly correlated to fibromatosis-like and spindle cell sarcoma molecular subtypes. • ZEB1 and Vimentin were independent risk factors for the outcomes and might serve as a therapeutic target for MBC patients. [ABSTRACT FROM AUTHOR]

MicroRNA-431 (miR-431) has been recognized as an oncogenic miRNA, being implicated in the initiation and development of human cancers. Recently, deregulation of miR-431 has been reported in several tumors. However, the clinical significance of miR-431 and its underlying role in human hepatocellular carcinoma (HCC) are poorly explored. Herein, we found that miR-431 expression was reduced in HCC tissues compared to noncancerous tissues. Otherwise, down-regulation of miR-431 was observed in aggressive tumor tissues. The levels of miR-431 expression in HCC cell lines were significantly lower than that in a nontransformed hepatic cell line. Clinical association analyses disclosed that a low level of miR-431 was prominently associated with poor prognostic features of HCC including venous infiltration, high Edmondson–Steiner grading and advanced tumor-node-metastasis (TNM) tumor stage. Our in vitro studies showed that up-regulation of miR-431 expression reduced cell invasion and migration in HCCLM3 cells. In contrast, down-regulation of miR-431 expression promoted SMMC-7721 cell invasion and migration. We found that up-regulation of miR-431 expression decreased zinc finger E-box binding homeobox 1 (ZEB1) expression and inhibited the epithelial–mesenchymal transition (EMT) with increased E-cadherin expression and decreased vimentin expression in HCCLM3 cells. Otherwise, down-regulation of miR-431 expression increased ZEB1 expression and promoted EMT in SMMC-7721 cells. Significantly, ZEB1 was identified as a target of miR-431 in HCC. ZEB1 knockdown abrogated the effect of miR-431 silencing on EMT and cell mobility in SMMC-7721 cells. In conclusion, miR-431 inhibits migration and invasion of HCC cells by suppressing ZEB1-mediated EMT. [ABSTRACT FROM AUTHOR]

The main aim of the study is to explore the mechanism of microrNA-145-3p targeting ZEB1 on migration and invasion in cervical cancer cells. In this study, cervical cancer cell line C33A was selected and transfected. After transfection of C33A cells, the expression level of Mir-145-3p was measured by QT-PCR. Human cervical cancer cell line (C33A) was cultured in RPMI 1640 medium and cell transfection experiment was performed. The related targets of Mir-145-3p, and a complementary binding site was found between the 3′UTR terminal of ZEB1 and Mir-145-3p was performed using a Targetscan biological software. The luciferase reporter system was used to determine the targeting relationship between Mir-145-3p and complementary binding site. The expression of e-cadherin and vimentin proteins was detected by western blot analysis. After 48 h, the expression level of Mir-145-3p in mimics group was significantly increased, the inhibitor group was significantly down-regulated and was statistically significant (P < 0.05). The clone formation, migration and invasion ability and vimentin expression in the MIMics group were significantly decreased than NC group. However, the apoptosis rate and e-cadherin protein expression were significantly increased than control group. In the inhibitor group, the cell cloning, cell migration, invasion and vimentin protein expression were significantly increased, while the cell apoptosis rate and e-cadherin protein expression were significantly decreased (P < 0.05). Analysis of luciferase expression genes revealed that mirNa-145-3p and ZEB1 have targeted regulatory relationship. Compared with Mir-NC, there was no significant difference between Mir-145-3p MUT and Mir-NC (P > 0.05), and luciferase activity of Mir-145-3p WT was significantly down-regulated (P > 0.05). Overexpression of microrNA-145-3p downregulated ZEB1 expression and inhibited proliferation, migration, invasion, and epithelial-mesenchymal transformation of cervical cancer cells, and promoted cell apoptosis. [ABSTRACT FROM AUTHOR]

Epithelial—mesenchymal transition (EMT), a critical step in the acquisition of metastatic state, is an attractive target for therapeutic interventions directed against tumor metastasis. Honokiol (HNK) is a natural phenolic compound isolated from an extract of seed cones from Magnolia grandiflora. Recent studies from our lab show that HNK impedes breast carcinogenesis. Here, we provide molecular evidence that HNK inhibits EMT in breast cancer cells resulting in significant downregulation of mesenchymal marker proteins and concurrent upregulation of epithelial markers. Experimental EMT induced by exposure to TGFβ and TNFα in spontaneously immortalized nontumorigenic human mammary epithelial cells is also completely reversed by HNK as evidenced by morphological as well as molecular changes. Investigating the downstream mediator(s) that may direct EMT inhibition by HNK, we found functional interactions between HNK, Stat3, and EMT‐signaling components. In vitro and in vivo analyses show that HNK inhibits Stat3 activation in breast cancer cells and tumors. Constitutive activation of Stat3 abrogates HNK‐mediated activation of epithelial markers whereas inhibition of Stat3 using small molecule inhibitor, Stattic, potentiates HNK‐mediated inhibition of EMT markers, invasion and migration of breast cancer cells. Mechanistically, HNK inhibits recruitment of Stat3 on mesenchymal transcription factor Zeb1 promoter resulting in decreased Zeb1 expression and nuclear translocation. We also discover that HNK increases E‐cadherin expression via Stat3‐mediated release of Zeb1 from E‐cadherin promoter. Collectively, this study reports that HNK effectively inhibits EMT in breast cancer cells and provide evidence for a previously unrecognized cross‐talk between HNK and Stat3/Zeb1/E‐cadherin axis. Highlights: Shows that honokiol inhibits EMT in breast cancer cells.Reveals that Stat3 inhibition is integral for honokiol‐mediated EMT inhibition.Mechanistic insights show the involvement of Stat3/Zeb1/E‐cadherin axis.Proposes that honokiol could potentially be a rational therapeutic strategy for breast cancer. [ABSTRACT FROM AUTHOR]

Growing evidence illustrates that aberrant activation of the epithelial-mesenchymal transition plays a key role in tumor cell invasion and metastasis. Transcription factors SNAI1 and ZEB1 regulate the epithelial-mesenchymal transition. To determine if SNAI1 and ZEB1 are involved in the metastasis of cervical cancer, we used immunohistochemistry to evaluate the expression of SNAI1, ZEB1, and vimentin in tumor and stromal compartments for a large set of cervical carcinoma samples. Results were evaluated using an H score (percentage × intensity). Of 70 samples, 64 cases (91.43%) were positive for SNAI1 expression. Themedian SNAI1H score was 174.00 (range, 5-285). Sixty-seven cases (95.71%)were positive for ZEB1, with a median H score of 165.77 (range, 5-260). Nuclear expression of SNAI1 and ZEB1 in tumor cells was positively associated with International Federation of Gynecology and Obstetrics (FIGO) stages (P = .015 and P = .008, respectively) and lymph node metastasis (P = .007 and P = .007, respectively); meanwhile, expression of vimentin in tumor cells was positively associated with lymph node metastasis (P = .019). According to negative vimentin expression, nuclear expression of ZEB1 in tumor cells was positively associated with FIGO stages (P = .04). According to positive vimentin expression, nuclear expression of SNAI1 in tumor cells was positively associated with FIGO stages (P = .018) and pN (P = .029). In light of these findings, we propose that SNAI1 and ZEB1 have the potential to be used as a novel predictor of pelvic lymph node metastasis and represent promising therapeutic targets in patients with cervical cancer. [ABSTRACT FROM AUTHOR]

Summary: Snail1 and ZEB1 are transcriptional repressors that drive tumor initiation and metastasis in animal models. Snail1 and ZEB1 are frequently coexpressed in tumor cell lines, suggesting that these factors may cooperate to promote tumor progression. However, coexpression of these transcriptional repressors in primary human cancer specimens has not been investigated. Previous studies assessed expression in primary breast cancers of Snail1 messenger RNA, which does not reflect Snail1 activity because Snail1 is subject to posttranslational modifications that inhibit its nuclear localization/activity. In the current study, using breast tumor cell lines of known Snail1 and ZEB1 expression status, we developed immunohistochemistry protocols for detecting nuclear Snail1 and nuclear ZEB1 proteins. Using these protocols, we assessed nuclear Snail1 and nuclear ZEB1 expressions in primary human breast cancers of varying subtypes (n = 78). Nuclear Snail1 and estrogen receptor α expressions were inversely associated in primary breast cancers, and nuclear Snail1 was expressed in approximately 80% of triple-negative breast cancers (lacking estrogen receptor α, progesterone receptor, and human epidermal growth factor receptor 2 overexpression). In contrast, nuclear ZEB1 was expressed at a significantly lower frequency in these breast cancers. Notably, nuclear Snail1 protein was detected in 45% of ductal carcinoma in situ specimens (n = 29), raising the important possibility that nuclear Snail1 expression in early stage breast lesions may predict future development of invasive breast cancer. Collectively, our studies demonstrate frequent expression of nuclear Snail1, but not nuclear ZEB1, in invasive, triple-negative breast cancers as well as in intraductal carcinomas. [Copyright &y& Elsevier]

Abstract: The p53 protein family consists of three transcription factors: p53, p63, and p73. These proteins share significant structural and functional similarities and each has unique biological functions as well. Although the role of p53 in cellular stress is extensively studied, many questions remain about p63 and p73. In this review we summarize current data on functional interactions within the p53 family, their regulation and roles in response to genotoxic stress. We also discuss the significance of p73 and p63 for cancer therapy and outline novel approaches in development of therapeutic drugs that specifically target the p53 family. [Copyright &y& Elsevier]