MicroRNA-29b-3p suppresses oral squamous cell carcinoma cell migration and invasion via IL32/AKT signalling pathway.
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ABSTRACT:
SUBMITTER: He J
PROVIDER: S-EPMC6933408 | biostudies-literature | 2020 Jan
REPOSITORIES: biostudies-literature
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Project description:OBJECTIVES:Tongue squamous cell carcinoma (TSCC) is the most frequent type of oral malignancy. Increasing evidence has shown that miRNAs play key roles in many biological processes such as cell development, invasion, proliferation, differentiation, metabolism, apoptosis and migration. MATERIALS AND METHODS:qRT-PCR analysis was performed to measure miR-137 expression. CCK-8 analysis, cell colony formation, wound-healing analysis and invasion were performed to detect resultant cell functions. The direct target of miR-137 was labelled and measured by luciferase assay and Western blotting. RESULTS:We demonstrated that expression of miR-137 was downregulated in TSCC tissues compared to matched normal ones. miR-137 expression was downregulated in TSCC lines (SCC4, SCC1, UM1 and Cal27) compared to the immortalized NOK16B cell line and normal oral keratinocytes in culture (NHOK). In addition, we have shown that miR-137 expression was epigenetically regulated in TSCCs. Overexpression of miR-137 suppressed TSCC proliferation and colony formation. Ectopic expression of miR-137 promoted expression of the epithelial biomarker, E-cadherin, and inhibited the mesenchymal biomarker, N-cadherin, as well as vimentin and Snail expression, indicating that miR-137 suppressed TSCC epithelial-mesenchymal transition (EMT). We also showed that ectopic expression of miR-137 inhibited TSCC invasion and migration. In addition, we identified SP1 as a direct target gene of miR-137 in SCC1 cells. SP1 overexpression rescued inhibitory effects exerted by miR-137 on cell proliferation and EMT. CONCLUSIONS:These results indicate that miR-137 acted as a tumour suppressor in TSCC by targeting SP1.
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Project description:The aim of the present study was to investigate the effects of microRNA (miR)‑29b‑3p gene promoter methylation on angiogenesis, invasion, and migration in human pancreatic cancer. Prediction of promoter methylation of miR‑29b‑3p was performed through the MethPrimer tool. Then the methylation levels of miR‑29b‑3p in human pancreatic cancer tissues and cell lines were detected by pyrosequencing, and the relative expression of miR‑29b‑3p was assessed in pancreatic cancer tissues by qPCR. The results were analyzed by linear regression. Western blot analysis was used to detect expression of DNA methyltransferases (DNMTs) in pancreatic cancer tissues and adjacent tissues. The Transwell assay was used to detect the ability of cell migration and invasion. Cells were co‑cultured with human umbilical vein endothelial cells (HUVECs) to detect the ability of angiogenesis. The results revealed that DNMT1 expression in pancreatic cancer tissues was higher than that in adjacent tissues. Further results showed that expression of miR‑29b was negatively correlated with the methylation level of the miR‑29b promoter. Bxpc3 and Capan‑2 cells had higher methylation levels, and the expression level of miR‑29b‑3p in Bxpc3 and Capan‑2 cells was found to be lower than that of other cell lines. Expression of zonula occludens‑1 (ZO‑1) and occludin was significantly increased, and the migration of cancer cells was decreased after cells were treated with siRNA DNMT1. Further results showed that miR‑29b reversed the promotive effect of DNMT1 overexpression on tumor cell malignant properties. Methylation of the miR‑29b‑3p promoter contributes to angiogenesis, invasion, and migration in pancreatic cancer. This study indicated that the alteration of methylation of mR‑19b may be a potential approach for inhibiting the progression of pancreatic cancer.
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Project description:MicroRNAs (miRNAs) have been shown to function as either oncogenes or tumor suppressors by negatively regulating target genes involved in tumor initiation and progression. In this study, we demonstrated that down-regulation of miR-33a-3p in human primary hepatocellular cancer (HCC) specimens was significantly associated with metastases and poor survival. Over-expression of miR-33a-3p in HepG2 cells remarkably suppressed not only cell growth, migration and invasion, but also tumor growth and metastases in the chick embryo chorioallantoic membrane (CAM) assay, and down-regulated Pre-B-Cell Leukemia Homeobox 3 (PBX3) expression. Conversely, inhibition of miR-33a-3p in Bel-7402 cells resulted in increased of cell growth, spreading and invasion. Furthermore, rescue experiments by over-expression PBX3 completely eliminated the inhibitory effects of miR-33a-3p on tumor growth and metastasis, both in vitro and in vivo. The luciferase assay showed that 3'-untranslated regions (3'-UTRs) of PBX3 were inhibited significantly by miR-33a-3p, while mutations in the miR-33a-3p pairing residues rescued the luciferase expression. Taken together, our findings suggest that miR-33a-3p suppressed the malignant phenotype while also inhibiting PBX3 expression in hepatocellular cancer, implying that miR-33a-3p may be a promising biomarkers and therapy target for HCC intervention.
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Project description:Invasion and metastasis are major contributors to treatment failure in patients with head and neck squamous cell carcinomas (HNSCCs) and microRNAs (miRNAs) are reported to play important roles in tumor progression. Our study therefore try to find the crucial miRNAs and reveal their molecular and functional mechanisms involved in migration and invasion of HNSCCs. Through The Cancer Genome Atlas (TCGA) data analysis, we screened out miR-3187-3p and its biological function and specific mechanism were further analyzed. The wound-healing and transwell invasion assay demonstrated that miR-3187-3p promoted the capacity of migration and invasion of HNSCCs in vitro. Luciferase reporter assays showed that PER2 was a direct target of miR-3187-3p, which could reverse the effect of miR-3187-3p in HNSCCs. Furthermore, we found that miR-3187-3p / PER2 axis activated the Wnt / β-catenin signaling pathway in HNSCCs. Altogether, our study indicated that miR-3187-3p enhanced migration and invasion by targeting PER2 in HNSCCs.
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