Effect of up-regulating the expression of microRNA-613 on biological behavior of human renal cell carcinoma cells / 中国医师杂志

ABSTRACT

Objective To observe the effect of microRNA-613 (miR-613) on the biological behavior of human renal cell carcinoma cells and explore its molecular mechanism.Methods The synthesized miR-613 mimics and miR-NC were respectively transferred into 786-0 cells by Lipofectamine 3000 and divided into experimental group and control group.Real time quantitative polymerase chain reaction (qPCR) was used to detect the expression of miR-613 in cells of two groups to verify the transfection success.The proliferation of 786-0 cells was detected by methyl thiazolyl tetrazolium (MTT) assay and the migration of 786-0 cells was detected by Transwell migration assay.Bioinformatics predicts the target gene of miR-613.Dual luciferase reporter assay validates the targeted relationship between miR-613 and target genes,and their binding sites.qPCR and Western blot were used to detect the expression of target genes.Results qPCR test results showed that the expression of miR-613 in the experimental group (16.22 ± 1.08) was significantly higher than that in the control group (1.06 ± 0.20),with statistically significant difference (P ＜ 0.01).The results of MTT showed that the cells in the experimental group showed a decrease in absorbance value at the d-th point after transfection (P ＜ 0.05).Transwell cell migration test results showed that the cells in the control group and experimental group migrated from the upper chamber of Transwell to the lower chamber of the cells were respectively (95.55 ± 17.88) and (199.10 ± 22.74),with statistically significant difference (P ＜ 0.05).Bioinformatics predicts that the target gene of miR-613 is Cortactin (CTTN).Dual luciferase reporter assay confirmed that miR-613 can target CTTN gene (P ＜ 0.05).miR-613 can significantly inhibit the CTTN gene expression in renal cell carcinoma cells (P ＜ 0.01).Conclusions Upregulation of miR-613 expression can inhibit the proliferation and migration of renal cell carcinoma cells,and its possible molecular mechanism is that miR-613 inhibits the expression of CTTN gene.