中文摘要：

目的探讨慢病毒介导LATS1过表达对人肾癌细胞786-0增殖、凋亡及下游癌基因YAP的影响。方法采用携带LATS1基因的慢病毒载体感染人透明细胞肾癌细胞系786-0,利用RT-PCR检测感染后786-0细胞中Hippo-YAP信号通路大肿瘤抑制基因-1（large tumor suppressor gene 1,LATS1）mRNA和下游癌基因Yes-相关蛋白（Yes-associated protein,YAP）mRNA的表达水平,Western blot检测LATS1和YAP的蛋白表达水平,流式细胞术（flow cytometry,FCM）检测细胞凋亡和周期,细胞增殖/毒性试剂（cell counting kit-8,CCK-8）检测细胞增殖抑制情况。结果慢病毒介导LATS1感染肾癌786-0细胞系后,LATS1 mRNA及蛋白表达水平显著高于对照组及空病毒组（P〈0.05）,YAP mRNA及蛋白表达水平显著低于对照组及空病毒组（P〈0.05）,细胞周期停滞在G1期（P〈0.05）、细胞增殖受到明显抑制（P〈0.05）、细胞凋亡显著增加（P〈0.05）。结论慢病毒介导LATS1过表达下调YAP基因表达,抑制786-0细胞增殖、促进其凋亡并阻滞细胞周期于G1期。肾癌的发生、发展可能与LATS1和YAP表达失调有关。

英文摘要：

Objective To investigate the effect of lentivirus-mediated large tumor suppressor gene 1（ LATS1） overexpression on the cell proliferation,apoptosis and downstream oncogene Yes-associated protein（ YAP） in human clear cell renal cell carcinoma（ ccRCC） cell line 786-0. Methods LATS1 gene was introduced into 786-0 cells via lentiviral vector. The mRNA expression levels of LATS1 and YAP in Hippo-YAP signaling pathway were detected by RT-PCR,while the protein levels were detected by Western blotting. The cell apoptosis and cell cycle were analyzed by flow cytometry（ FCM）,and the cell proliferation inhibition was assayed by cell proliferation and toxicity detection reagent（ cell counting kit-8）. Results After 786-0 cells were transfected with LATS1 gene,the expression of LATS1 at mRNA and protein levels was significantly increased（ P 0. 05）,while the expression of YAP at mRNA and protein levels was markedly decreased（ P 0. 05）. After lentiviral-mediated LATS1 overexpression,the cell cycle was arrested at G1phase（ P 0. 05）,the cell proliferation was obviously inhibited（ P 0. 05）,and the cell apoptosis was increased significantly（ P 0. 05）. Conclusion Lentivirus-mediated LATS1 overexpression down-regulates the expression of YAP,inhibits the cell proliferation of 786-0 cells,induces the cell apoptosis and blocks the cell cycle at G1 phase.The occurrence and development of RCC may be associated with expression disorder of LATS1 and YAP.