Effects of lentivirus-mediated shRNA silencing YAP gene on proliferation，apoptosis，and androgen receptor of human prostate cancer cell line LNCaP cells

Abstract:

Objective：To explore the effect of lentivirus-mediated shRNA silencing Yes-associated protein（YAP） gene on prolifera－tion，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（real-time PCR） and Western blot were used to detect mRNA and protein ex－pression 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. Apoptosis of LNCaP cells was examined by flow cytometry（FCM）. Results：YAP-shRNA lentiviral interference vectors were suc－cessfully infected into LNCaP cells. The infection efficiency of lentiviral interference vectors was more than 95% after puro-mycin selection and the stable YAP knockdown in LNCaP cells was established. Compared with those in control group and negative control group，expressions of YAP mRNA and protein were significantly reduced after YAP-shRNA lentiviral interference vectors being infected（P=0.00），meanwhile expressions of AR mRNA and protein were also significantly down-regulated（P=0.000）；in YAP-shRNA lentiviral interference vectors group，cell proliferation was inhibited，and apoptosis was increased significantly（P=0.000）. Conclusions：lentivirus-mediated shRNA silencing YAP gene can down-regulate the expression of YAP gene，inhibit the proliferation and promote apoptosis in LNCaP. YAP may act as a ligand of AR and regulate androgen receptor signaling pathway.