Study On The Effects Of MicroRNA-183 On The Proliferation, Apoptosis, Migration, Invasion And The Expression Of EGR-1, PTK6 Of HepG2 Cells

Objective: To study the effects of miR-183 on proliferation,apoptosis,migration,invasion of human hepatoma HepG2 cells,and to investigate its effects on early growth response factor 1(EGR-1)and protein tyrosine kinase 6(PTK6),in attempt to provide theoretical foundations for targeted therapy of liver cancer.Methods: Human hepatoma HepG2 cells were routinely cultured.The liposome transfection method was used to interfere with miR-183 expression in HepG2 cells.The experiment was divided into three groups: Control group(without any treatment),negative control(NC)group(transfected with the negative control sequence)and miR-183 inhibitor group(transfected with miR-183 inhibitor).The proliferation ability of HepG2 cells was examined by MTT assay.The apoptosis ability was assessed by flow cytometry.The lateral migration ability of HepG2 cells in each group was observed by wound healing assay.Longitudinal migration and invasion ability of HepG2 cells were detected by Transwell migration and invasion assay.Real-time fluorescence quantitative PCR was used to detect the expression level of miR-183,EGR-1 and PTK6 in HepG2 cells after transfection;The protein levels of EGR-1 and PTK6 in HepG2 cells in each group were detected by Western blotting.Bioinformatics software were used to predict the targeting regulatory relationship between miR-183 and EGR-1.Result: The result of liposome transfection showed that the miR-183 expression levels in HepG2 cells of miR-183 inhibitor group were significantly lower than that in Control group and NC group(P<0.01).MTT experimental result suggested that the proliferation rate of HepG2 cells in mi R-183 inhibitor group was significantly lower than that of Control group and NC group(P<0.01).Flow cytometry results showed that the apoptosis rate of HepG2 cells in mi R-183 inhibitor group was significantly higher than that in Control group and NC group(P<0.01).The result of wound healing assay showed that a significant decrease of wound healing rate in miR-183 inhibitor group than Control group and NC group(P<0.01).Transwell assay results showed that the number of transmembrane cells in the miR-183 inhibitor group was significantly lower than that in Control group and NC group(P<0.01).Real-time fluorescence quantitative PCR results showed that the levels of EGR-1 gene in miR-183 inhibitor group was significantly higher than that in Control group and NC group(P<0.01),and the levels of PTK6 gene was significantly lower in Control group and NC group(P<0.01).Western blotting result showed that the ptotein expression levels of EGR-1 in miR-183 inhibitor group were significantly increased(P<0.01),and the gene expression levels of PTK6 were significantly decreased(P<0.01).Bioinformatics result show that miR-183 targets the EGR-1 gene.Conclusion:?miR-183 promotes the proliferation,migration,and invasion of human hepatoma HepG2 cells and inhibits apoptosis.?In HepG2 cells,EGR-1 is a potential target gene of miR-183.?In HepG2 cells,mi R-183 indirectly affected the expression levels of PTK6.