Influence of YAP gene silencing by shRNA on the proliferation and apoptosis of human renal cancer cell 786-O

Abstract:

Objective：To investigate the effect of Yes-associated protein（YAP） gene silencing by shRNA on the proliferation and apoptosis of human renal cancer cells 786-O. Methods：Expression of YAP protein in 786-O cells was detected by immunofluores－cence assay. Lentiviral interference vectors of shRNA-YAP were constructed successfully and were transfected into 786-O cells. Meanwhile blank control group and negative control group were divided. Expression of YAP mRNA and protein in 786-O cells was detected by RT-PCR and Western blot，respectively. CCK-8（cell counting kit-8） assay was applied to examine the effect of YAP si－lencing on the proliferation in 786-O cells. Apoptosis and cell cycle of 786-O cells were examined by flow cytometry. Results：YAP protein was expressed in cytoplasm and nucleus of 786-O cells. On 4 d after shRNA-YAP lentiviral interference vectors being in－fected，expression of YAP mRNA and protein in 786-O cells was significantly reduced（P=0.000），cell proliferation was inhibited，and apoptosis was increased significantly（P=0.000）. YAP silencing resulted in cell cycle distribution disorder in 786-O cells，which the number of G1 phase cells was significantly increased while the number of S phase cells was significantly decreased compared with that of blank control group and negative control group（P=0.000）. Conclusion：shRNA-YAP lentiviral interference vectors can effectively decrease YAP gene expression in 786-O cells，inhibit cell proliferation and promote cell apoptosis.