Yes相关蛋白在肝癌上皮间质转化中的作用  

Silence Yes-associated protein expression inhibit the invasion ability of hepatocellular carcinoma cells

作　　者：王寿华[1] 李华[1] 张彤[1] 吴小材[1] 乔欣[1] 陈规划[1] 

机构地区：[1]中山大学附属第三医院肝移植中心中山大学器官移植研究所广东省器官移植研究中心,广州510630

出　　处：《中华肝胆外科杂志》2014年第11期805-809,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：国家重点基础研究发展计划（973计划）（2009CB522404）;国家自然科学基金（81172038）

摘　　要：目的 探讨Yes相关蛋白（YAP）表达在肝癌上皮间质转化（EMT）中的作用.方法 通过YAPsiRNA（h）转染肝癌细胞MHCC97H及MHCC97L沉默YAP表达,通过实时PCR、蛋白印迹检测实验组与对照组肝癌细胞EMT标志物E-cadherin、N-cadherin表达变化,Transwell侵袭实验检测细胞侵袭能力的变化.结果 YAP siRNA（h）沉默肝癌细胞YAP蛋白72 h后与对照组比较,在高侵袭性MHCC97H细胞,实验组E-cadherin蛋白表达升高明显（P＜0.05）,N-cadherin蛋白表达显著下降（P＜0.05）.在低侵袭性MHCC97L细胞,实验组E-cadherin蛋白升高明显（P＜0.05）,N-cadherin蛋白无明显下降（P＞0.05）.Transwell侵袭实验结果显示,与对照组穿孔细胞数比较,在高侵袭性MHCC97H细胞,实验组穿孔细胞显著减少[（66±6.89）比（117±7.23）,P＜0.05].与对照组穿孔细胞数比较,在低侵袭性MHCC97L细胞,实验组穿孔细胞减少明显[（40±2.65）比（77±4.33）,P＜0.05].实时PCR检测两种肝癌细胞E-cadherin mRNA表达水平有所升高但不明显（P＜0.05）,而N-cadherin mRNA表达水平则无明显变化（P＞0.05）,考虑E-cadherin、N-cadherin蛋白表达变化可能存在转录后调控机制.结论 YAP蛋白表达与肝癌细胞上皮间质转化（EMT）存在相关性,抑制YAP蛋白的表达能够显著降低肝癌细胞的侵袭能力.Objective To investigate the correlation between the Yes-associated protein （YAP） expression and epithelial-mesenchymal transition （EMT） by transiently transfecting with YAP small-interfering RNA （siRNA） in MHCC97H and MHCC97L cells.Methods MHCC97H and MHCC97L cells were transiently transfected by YAP siRNA.Furthermore,protein expressions and mRNA levels of characteristic markers of EMT （E-cadherin,N-cadherin） were examined by Western blotting and real-time polymerase chain reaction.Transwell invasion assay was used to detect changes of invasiveness of MHCC97H and MHCC97L cells.Results The YAP siRNA transfected group in MHCC97H was examined after 72 hours by Western blotting.The result showed obviously higher expression of E-cadherin in transfected group 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 group cells significantly decreased in MHCC97H（66 ± 6.89 vs 117 ± 7.23,P ＜ 0.05）,and compared with the control group,the number of the transfected group also significantly decreased in MHCC97L （40 ±2.65 vs 77 ±4.33,P ＜0.05）.The result of real-time polymerase chain reaction indicated that mRNA levels of E-cadherin increased （P ＜ 0.05）,but mRNA levels of N-cadherin did not significantly change （P ＞ 0.05）.This is considered as post-transcriptional regulation after silencing YAP in MHCC97H and MHCC97L.Conclusions YAP silencing is able to inhibit EMT in MHCC97H and MHCC97L cells by modulating the characteristic markers of EMT.The inhibition of YAP expression can reduce the invasion ability of hepatocellular carcinoma cells.

关 键 词：肝细胞癌 Yes相关蛋白 上皮间质转化 

分 类 号：R735.7[医药卫生—肿瘤]