Effects Of Lentivirus-mediated YAP Gene Silencing On Proliferation And Apoptosis Of Renal Carcinoma Cells

Objective: To investigate the effect of Yes-associated protein (YAP)gene silencing by shRNA lentiviral interference vectors on proliferation andapoptosis of human renal cancer cells786-0.Methods：786-0renal carcinoma cells were cultured in RPMI1640medium. To construct shRNA lentiviral interference vector targeted YAPgene and then transfected into786-0cells, meanwhile setting blank controlgroup and negative control group. The mRNA and protein expression levelof YAP and the downstream transcription factor TEAD1in786-0cells weredetected by RT-PCR and Western blot, respectively. CCK-8(cell countingkit-8) assay was applied to examine the effect of YAP silencing on theproliferation in786-0cells. Apoptosis and cell cycle of786-0cells wereexamined by flow cytometry (FCM).Results: Three specific YAP-shRNA lentiviral vectors and negativecontrol shRNA lentiviral vectors were constructed and transfected into786-0cells successfully. After YAP-shRNA lentiviral interference vectorsinfected786-0cells96hours, expression of YAP mRNA and protein in 786-0cells was significantly reduced (P=0.000), and TEAD1mRNA andprotein expression levels were also significantly decreased (P=0.000).shYAP-3transfected group cells proliferation was inhibited, and apoptosisincreased significantly (P=0.000), YAP silencing resulted in cell cycledistribution disorder in786-0cells, which the number of G1phase cells wassignificantly increased comparing with blank control group and negativecontrol group, while the number of S phase cells was significantly decreased(P=0.000).Conclusion: shRNA-YAP lentiviral interference vectors caneffectively decrease YAP genes expression in786-0cells, inhibit cellproliferation and promote cell apoptosis.