YAP蛋白在结肠癌中的表达及其对SW480细胞增殖和侵袭的影响

摘要

目的:研究YAP蛋白在结肠癌中的表达及临床意义,并初步探讨其对结肠癌细胞增殖、侵袭的影响.方法:免疫组化检测YAP蛋白在25例结肠癌组织及配对正常组织中的表达,并分析其与结肠癌临床病理特征之间的关系;用Real-Time PCR和 Western blot验证YAP-shRNA慢病毒载体转染效率;生长曲线实验和MTT法检测下调YAP表达对结肠癌细胞SW480增殖的影响;Transwell实验检测下调YAP表达对结肠癌细胞SW480侵袭的影响;Western blot检测下调YAP表达对CyclinD1、E-cadherin及Vimentin蛋白的影响.结果:YAP在结肠癌组织中的表达显著高于正常结肠组织(P＜0.05),并且与淋巴结转移密切相关(P＜0.05);转染YAP-shRNA慢病毒载体的SW480细胞YAP的表达在mRNA和蛋白水平明显降低(均P＜0.05);下调YAP表达后结肠癌细胞增殖及侵袭明显减弱(均P＜0.05);下调YAP表达可抑制结肠癌细胞CyclinD1、E-cadherin及Vimentin蛋白的表达(均P＜0.05).结论:YAP在结肠癌组织中高表达,且与淋巴结转移相关,下调YAP表达可抑制结肠癌细胞增殖及侵袭能力,预示YAP在结肠癌恶性进展中具有重要的作用.%Objective:To investigate the expression of YAP in colon cancer tissues and role of YAP on the prolif-erative and invasive ability of colon cancer cells.Methods:The expression of YAP in 25 colon cancer tissues and paired normal tissues was determined by immunohistochemical staining.The relationship between the expression of YAP and its clinicopathologic features was further analyzed.Human colon cancer cells SW480 were transfected with the YAP－shRNA lentiviral vectors,and the YAP mRNA and protein expressionle were determined by Real－Time PCR and Western blot analysis.The effect of down－regulation of YAP expression on the proliferative and invasive a-bility was examined by cell growth curve assay,MTT assay and Transwell assay,respectively.Protein levels of Cy-clinD1,E－cadherin and Vimentin were detected by Western blot analysis.Results:The expression of YAP was sig-nificantly increased in colon cancer tissues,compared to the normal tissues.YAP expression was associated with lymph node metastasis.In SW480 cells transfected by YAP－shRNA lentiviral vectors,the mRNA and protein expression lev-els of YAP mRNA and protein were significantly down－regulated(both P＜0.05).Down－regulation of YAP expres-sion inhibited proliferative and invasive ability of SW480 cells(both P＜0.05).The protein levels of CyclinD1,E－cadherin and Vimentin were decreased in YAP－shRNA SW480 cells(all P＜0.05).Conclusion:The expression of YAP in colon cancer tissues was associated with lymph node metastasis,and YAP plays an important role on prolifera-tive and invasive ability of colon cancer cells.