Effects of miR-143 on proliferation and apoptosis of renal cell carcinoma GRC-1 cells and its target HIF-1α

LI Dong; LIU Xisheng; WANG Xinming; WANG Shuai; TAO Xiaoxin

Abstract：Objective To investigate the effects of miR-143 on proliferation and apoptosis of renal cell carcinoma GRC-1 cells and its target hypoxia inducible factor-1α( HIF-1α) . Methods Renal cell carcinoma cell line GRC-1 cells were transfected with miR-143 mimics, and real time quantitative polymerase chain reaction ( qPCR) was performed to evaluate the efficiency of transfection. According to the experiment, the GRC-1 cells were divided into 3 groups:non-transfection group, miR-143 control group and miR-143 transfection group. MTT method was used to detect the proliferation of cells in each group. Flow cytometry was used to detect the apop-tosis of each group. The qPCR and Western blotting were employed to measure the mRNA and protein level of HIF-1α. Dual luciferase reporter gene was applied to verify the relationship between miR-143 and HIF-1α. Results The level of miR-143 in miR-143 transfec-tion group was higher than those in non-transfection group and miR-143 control group, and the difference was statistically significant ( P<0. 05) . Compared with other two groups, there were decreased proliferation rates but increased apoptotic rates in miR-143 transfection group ( P<0. 05) . Transient overexpression of miR-143 in GRC-1 cells decreased the expression of HIF-1αon both mRNA and protein levels. Moreover, dual-luciferase reporter assay confirmed that HIF-1α was a direct target of miR-143 in GRC-1 cells. Conclusion miR-143 may regulate the proliferation of GRC-1 cells, and HIF-1α is a direct target of miR-143.