Effect of exogenous small RNA transfection on the growth of renal clear cell carcinoma cells / 实用医学杂志

ABSTRACT

Objective To investigate the effect of exogenous small RNA (dsP21-397) transfection on growth of human renal clear cell carcinoma cell lines A-498 and Caki-1. Methods The dsControl(control group) and dsP21-397(experimental group)were transfected into A-498 and Caki-1,respectively. The expression of p21 mRNA was analyzed by qRT-PCR. The expression of p21 protein,CDK4 and Cyclin D1 proteins were detected by Western blot. The cell cycle distribution was examined by flow cytometry(FCM). MTS assay and colony formation assay were used to analyze cell viability and proliferation ability. Results Compared with dsControl,p21 mRNA levels in A-498 and Caki-1 cells increased to 2.55-fold(P<0.01)and 2.18-fold(P<0.01),respectively,after transfection with dsP21-397. The expression of p21 protein was up-regulated while the expression of CDK4 and CyclinD1 were down-regulated. The percentage of cells in G0/G1 phase significantly increased after transfection of dsP21-397,and the proportion of cells in S phase and G2/M phase significantly decreased. The activity of A-498 and Caki-1 cells significantly decreased and the number of colonies in the dsP21-397 group was significantly lower. Conclusions dsP21-397 can significantly activate p21 protein expression,down-regulate the cell cycle-associated proteins,and inhibit the growth of renal clear cell carcinoma cells.