Yes-associated protein (YAP) expression is involved in epithelial-mesenchymal transition in hepatocellular carcinoma
Yes-associated protein (YAP) expression is involved in epithelial-mesenchymal transition in hepatocellular carcinoma
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DOI：
10.1007/s12094-015-1353-4
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发表时间：
2016-02-01
影响因子：
3.4
通讯作者：
Chen, G.
中科院分区：
文献类型：
--
作者：
Wang, S.;Li, H.;Chen, G.
目的探讨RNA干扰下调yes相关蛋白(YAP)对MHCC97H和MHCC97L上皮间质转化过程的生物学影响。用YAP-siRNA瞬时转染MHCC97H和MHCC97L细胞。并采用Western blotting和实时聚合酶链反应检测上皮间质转化特征标志物（E-cadherin、N-cadherin）的蛋白表达及mRNA水平，并采用Transwell侵袭实验检测MHCC97H和MHCC97L细胞侵袭能力的变化。转染YAP-siRNA组72 h后Western blotting检测MHCC97H细胞侵袭能力的变化。 印迹结果显示，E-cadherin 表达较对照组明显升高（P < 0.05），N-cadherin 表达较低（P < 0.05）。在MHCC97L细胞中，E-cadherin的表达也显着升高（P < 0.05）；然而，N-cadherin 表达没有显着变化（P > 0.05）。此外，与对照组相比，Transwell侵袭实验显示，MHCC97H和MHCC97L细胞系中转染组的数量显着减少（均P < 0.05）。实时聚合酶链式反应结果表明，这两种细胞系中E-cadherin mRNA水平升高（P < 0.05），但N-cadherin mRNA水平没有明显变化（P > 0.05），表明沉默YAP后转录后调控机制产生了一定的影响。人肝癌细胞系MHCC97H和MHCC97L中YAP的表达与 上皮-间质转化的特征标志物、N-钙粘蛋白和E-钙粘蛋白表达。
To investigate biological impact of the downregulation of yes-associated protein (YAP) through RNA interference in the process of epithelial-mesenchymal transition in MHCC97H and MHCC97L.MHCC97H and MHCC97L cells were transiently transfected by YAP-siRNA. Furthermore, protein expressions and mRNA levels of characteristic markers of epithelial-mesenchymal transition (E-cadherin, N-cadherin) were examined by Western blotting and real-time polymerase chain reaction, and transwell invasion assay was used to detect changes of invasiveness of MHCC97H and MHCC97L cells.The transfected group with YAP-siRNA in MHCC97H after 72 h by Western blotting showed obviously higher expression of E-cadherin compared with the control group (P < 0.05), and lower expression of N-cadherin (P < 0.05). In MHCC97L cells, the expression of E-cadherin was also significantly increased (P < 0.05); however, N-cadherin expression did not significantly change (P > 0.05). Moreover, compared with the control group, Transwell invasion assay showed that the number of the transfected groups was significantly decreased in MHCC97H and MHCC97L cell lines (both P < 0.05). The result of real-time polymerase chain reaction indicated that mRNA levels of E-cadherin increased (P < 0.05), but the mRNA levels of N-cadherin did not significantly change (P > 0.05) in these two cell lines, indicating some effects of post-transcriptional regulation mechanism after silencing YAP.YAP expression in human hepatocellular carcinoma cell lines MHCC97H and MHCC97L is closely related with the characteristic markers of epithelial-mesenchymal transition, N-cadherin and E-cadherin expression.