PO-167 MicroRNA- 200 c, a tumour suppressor that modulate the expression of cancer stem cells markers and epithelial-mesenchymal transition in colorectal cancer

Abstract

IntroductionColorectal cancer (CRC) is the most frequently diagnosed cancer and the most common gastrointestinal cancer worldwide. Due to the presence of populations of ST cells that cause recurrence, the possibility of cancer treatment is very low, The epithelial-mesenchymal transition (EMT) and and subsequent metastasis by giving rise to new tumours. Wnt/β-catenin pathway play an important role in maintaining proliferation, invasion and metastasis of CSCs. miRNA-200c function as a tumor-suppressor gene and negatively regulate EMT-associated genes in cancer cells. The aim of current study was to evaluate the inhibitory role of miR-200c on EMT, β-catenin and CSCs markers in HCT-116 and SW48 cell lines.Material and methodsThe expression of miR-200c, EMT-related genes, β-catenin and CSCs markers were quantified by qRT-PCR. Further, expression of β-catenin and EMT-related proteins, invasion and migration were analysed by Western blot, invasion and migration assay kit, respectively. Spheroid formation assay was used to enrich colorectal CSCs from colorectal cancer cell lines (HCT-116 and SW48).Results and discussionsLNA-anti-miR-200c suppressed the endogenous miR-200c in transfected cells compared with the control. QRT PCR and Western blot analysis of LNA-anti-miR-200c transfected cells revealed a considerable increase in CSCs markers, Vimentin ZEB-1, N-cadherin and β-catenin expression, with a concomitant reduction in E-cadherin expression level. Migration and invasion of HCT-116 cells increased in transfected cells.ConclusionThe results of current study revealed that downregulation of miR-200c may be an important factor for the overexpression of CSCs markers and EMT related genes in CRC.