Overexpression of miR146a-5p, as a novel prognostic marker, inhibits migration of human HT-29 colorectal cancer cells via down-regulating metastasis-related genes expression

Introduction & Aim: microRNAs are a group of post-transcription regulators whose expressions are altered in various cancers and contribute to cancer progression. According to studies, the expression of miR-146a-5p in some cancers, including colorectal cancer, is reduced. The purpose of this study was to restore of microRNA-146a-5p in HT-29 cell line and investigate the expression of metastasis-related genes in these cells. Methods. In this study, miR-146a-5p mimic was transfected into HT-29 cells via JetPEI transfection reagent. Transfection efficiency was evaluated by flow-cytometry. To evaluate the effects of miR-146a-5p replacement on the inhibition of cell migration, wound healing assay was performed. Finally, the expression levels of miR-146a-5p, vimentin, MMP9 and C-Myc were measured by the qRT-PCR method in HT-29 cells. Results. According to the obtained data, results of wound healing assays showed that miR-146a-5p mimic inhibited cell migration in HT-29 cell line compared with control groups. Finally, data of gene expression showed that miR-146a-5p mimic reduced vimentin, MMP9 and C-Myc gene expression compared to control cells. Conclusion. According to this study, the activation metastatic pathways were due to the down regulation of miR-146a-5p. According to this, miR-146a-5p can inhibit cells migration, in vitro, through down-regulating the expression of metastasis-related genes. Hence, miR-146a-5p can be a new diagnostic biomarker and new therapeutic target for colorectal cancer.

Keywords:

Colorectal cancer , Metastasis , HT-29 cell line , microRNA-146a-5p