Effects Of Lentivirus-Mediated SHRNA Silencing YAP Gene On Proliferation, Apoptosis, And Androgen Receptor Of Human Prostate Cancer Cell Line Lncap Cells

Objective:To explore the effect of lentivirus-mediated shRNA silencing Yes-associated protein (YAP) gene on proliferation, apoptosis, and androgen receptor of hormone dependent human prostate cancer LNCap cells.Methods:Three shRNA for YAP gene lentiviral interference vectors were infected into LNCaP cells, then the LNCaP cells stably knockdown YAP gene expression were screened by puromycin. Real-time quantitative PCR (qPCR) and Western blot were used to detect mRNA and protein expression of YAP and androgen receptor(AR) after lentiviral interference vectors being infected respectively. shRNA lentiviral vector with the best silence efficiency was acted as experiment group. CCK-8 (cell counting kit-8) assay and colony formation were applied to evaluate the cell proliferation in LNCaP cells. Morphological changes of apoptosis was detected by Hoechst 33258 in LNCaP cells. Cell cycle and Apoptosis of LNCaP cells was examined by flow cytometry (FCM), the expression of p21.. Cyclin D1、Bcl-2 and Bax proteins were also detected by Western blot.Results:YAP-shRNA lentiviral interference vectors were successfully infected into LNCaP cells. The infection efficiency of lentiviral interference vectors was more than 95%after puromycin selection and the stable YAP knockdown in LNCaP cells was established. Compared with those in control group and negative control group, expression of YAP mRNA and protein were significantly reduced after YAP-shRNA lentiviral interference vectors being infected, meanwhile expression of AR mRNA and protein were also significantly down-regulated; in YAP-shRNA lentiviral interference vectors group,cell proliferation was inhibited, cell cycle was arrested in G1 phase, apoptosis was increased significantly, the expression of p21 and Bax protein were up-regulated, CyclinDl and Bcl-2 were down-regulated.Conclusions:lentivirus-mediated shRNA silencing YAP gene can down-regulate the expression of YAP gene, inhibit the proliferation and promote apoptosis in LNCaP, may regulate p21 and CyclinD1 to arrest in G1 phase, affect the expression of Bcl-2 and Bax induced cell apoptosis. YAP may act as a ligand of AR and regulate androgen receptor signaling pathway.