Effect of siRNA targeting RPL6 gene on the proliferation and apoptosis of human cervical cancer HeLa cells

YAN Feng; XI Yanni

Abstract：Objective To explore the effect of siRNA targeting RPL6 gene on the proliferation and apoptosis of human cervi?cal cancer HeLa cells. Methods After the optimization of the transfection conditions, 2 siRNA vector fragments ( siRPL6?1, siRPL6?2) targeting RPL6 gene were effectively transfected into HeLa cells of human cervical cancer(inhibition group), respectively. Mean?while, there were also empty transfection group whose cells were transfected with the antisense sequence ( siControl) and control group without any treatment. The RPL6 expression levels were detected by real?time quantitative PCR ( qRT?PCR) at 24, 48, 72 and 96 h after transfection, and the siRPL6 vector with the higher inhibition rate was used for the following experiments. Thiazolyl blue ( MTT) was used to detect the proliferation inhibition rate at 24, 48, 72, 96 h after transfection. The apoptosis and cell cycle of the cells were detected by flow cytometry after 48 and 96 h. The expressions of CDK2, cyclin A and p21CIP1 after 96 h were detected by Western blot?ting. Results The level of mRNA RPL6 in the inhibition group was lower than that in the empty transfection group and the control group ( P<0?05) . Since the interference efficiency of the siRPL6?2 fragment was higher than that of the siRPL6?1, so the subsequent trials chose the siRPL6?2 fragment. Compared with other two groups, the proliferation inhibitory rate, apoptosis rate and caspase?3 acti?vation rate together with the proportion of cells in G0/G1 phase increased, but the proportion of cells in S phase and G2/M phase de?creased in inhibition group with statistically significant difference ( P<0?05);After transfection, the levels of p21CIP1 increased but the levels of CDK2 and cyclin A were decreased in inhibition group versus other groups ( P<0?05) . Conclusion The inhibition of RPL6 gene expression by siRNA can inhibit the proliferation and induce apoptosis and cell cycle arrest, and it can be valuable for the preven?tion and treatment of cervical cancer.