Cao Jian-Jia, Zhao Xiu-Min, Wang De-Lin, Chen Ke-Hong, Sheng Xia, Li Wen-Bin, Li Mei-Cai, Liu Wu-Jiang, He Jiang
Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China.
Institute of Urology, First Hospital of Peking University, Beijing, P.R. China.
Oncol Rep. 2014 Oct;32(4):1594-600. doi: 10.3892/or.2014.3349. Epub 2014 Jul 22.
Yes-associated protein (YAP) has been reported to be an oncogene in a number of malignancies. It constitutes an important regulatory mechanism for the Hippo pathway, a key regulator of cell growth and apoptosis. The present study aimed to investigate the clinical significance and the role of YAP in the development of clear cell renal cell carcinoma (ccRCC). YAP expression levels were compared between ccRCC and adjacent normal renal tissues by RT-PCR and immunohistochemistry, respectively. YAP expression levels were then detected in ccRCC cell lines 786-0 and ACHN, as well as in human embryonic kidney 293 cells (HEK-293) using western blotting. Three specific YAP-shRNA lentiviral vectors were constructed and transfected into 786-0 cells, and then the mRNA and protein levels of YAP and downstream transcription factor TEAD1 were detected. Finally, the effects of YAP silencing on proliferation and the cell cycle distribution of 786-0 cells were detected by Cell Counting Kit-8 (CCK-8) and flow cytometry (FCM), respectively. The apoptosis rate was also analyzed by FCM. It was observed that the expression levels of YAP mRNA and protein in ccRCC tissues were higher than these levels in the adjacent normal renal tissues. The expression of YAP protein in ccRCC tissues was significantly correlated with clinical stage and differentiation. The YAP protein levels in the two ccRCC cell lines 786-0 and ACHN were significantly higher than that in the HEK-293 cells. Additionally, treatment of 786-0 cells with YAP-shRNA lentiviral vectors significantly reduced the expression levels of YAP and TEAD1 mRNA and protein. Further analyses in 786-0 cells in which YAP was decreased, revealed that cell proliferation was inhibited, cell cycle was arrested at the G1 phase and apoptosis was increased. These results indicate that YAP is an underlying oncogene in ccRCC and it may be a promising biomarker and therapeutic target of ccRCC.
Yes相关蛋白（YAP）在多种恶性肿瘤中被报道为一种癌基因。它构成了Hippo信号通路的重要调控机制，而Hippo信号通路是细胞生长和凋亡的关键调节因子。本研究旨在探讨YAP在透明细胞肾细胞癌（ccRCC）发生发展中的临床意义及作用。分别通过逆转录聚合酶链反应（RT-PCR）和免疫组织化学方法比较ccRCC组织与相邻正常肾组织中YAP的表达水平。然后利用蛋白质免疫印迹法检测ccRCC细胞系786-0和ACHN以及人胚肾293细胞（HEK-293）中YAP的表达水平。构建三种特异性YAP-shRNA慢病毒载体并转染至786-0细胞中，随后检测YAP及下游转录因子TEAD1的mRNA和蛋白水平。最后，分别通过细胞计数试剂盒-8（CCK-8）和流式细胞术（FCM）检测YAP沉默对786-0细胞增殖和细胞周期分布的影响。同时也通过FCM分析细胞凋亡率。结果发现，ccRCC组织中YAP mRNA和蛋白的表达水平高于相邻正常肾组织。ccRCC组织中YAP蛋白的表达与临床分期和分化程度显著相关。ccRCC细胞系786-0和ACHN中YAP蛋白水平显著高于HEK-293细胞。此外，用YAP-shRNA慢病毒载体处理786-0细胞可显著降低YAP和TEAD1 mRNA及蛋白的表达水平。对YAP表达降低的786-0细胞进行进一步分析发现，细胞增殖受到抑制，细胞周期停滞在G1期，细胞凋亡增加。这些结果表明，YAP是ccRCC中的潜在癌基因，可能是ccRCC有前景的生物标志物和治疗靶点。
