Effect of microRNA-1180 transfection on growth of renal cell carcinoma / 重庆医学

ABSTRACT

Objective To investigate the effect of microRNA-1180 transfection on the growth of renal cell carcinoma lines 786-O and ACHN.Methods The renal carcinoma cells were divided into the two groupscontrol group (transfecting dsControl) and experimental group (transfecting miR-1180).The expression change of p21 mRNA was detected by qRT-PCR.Western blot was conducted to analyze the expression changes of p21,CDK4,CDK6 and CyclinD1 proteins.Flow cytometry (FCM) was used to detect the cell cycle change.The MTS assay was conducted to detect the cell viability and the colony forming assay was performed to examine the cell proliferation ability.Results The qRT-PCR results showed that compared with the negative control dsControl group,after miR-1180 transfection,the expression level of·p21 mRNA in 786-O and ACHN cells was up-regulated to 2.54-fold and 2.49-fold respectively(P＜0.01).The expression trend of p21 protein was consistent with qRT-PCR results.The expression of CDK4,CDK6 and CyclinD1 proteins were significantly down-regulated.The FCM results showed that the proportion of cells in G0/ G1 phase was significantly increased after transfection of miR-1180,but the proportion of cells in S phase and G2/M phase was decreased significantly,indicating that the cell cycle was arrested in G0/G1 phase.The MTS assay results showed that compared with the dsControl group,the viability of the two kinds of renal carcinoma cells was significantly decreased.The colony formation assay showed that the number of colonies formed in the miR-1180 group was smaller,indicating the proliferation ability of miR-1180 transfected cells was decreased.Conclusion miR-1180 can significantly activate the p21 protein expression and inhibit the growth of renal carcinoma cell lines 786-O and ACHN.