siRNA干扰技术沉默p66SHC对结直肠癌细胞生物学特性的影响  

Effect of silencing p66SHC by siRNA on biological characteristics of colorectal cancer cells

作　　者：杨洲[1] 刚海菊[2] 覃先蓬[1] 贾贵清[1] 王波[1] 杨春[1] YANG Zhou;GANG Hai-ju;QIN Xian-peng(1.Department of Gastrointestinal Surgery,Sichuan Provincial People＇s Hospital,Sichuan Academy of Medical Sciences;Chengdu Vocational and Technical College Medical Branch Nursing Teaching and Research Office; Chengdu Sichuan 610041, China)

机构地区：[1]四川省医学科学院.四川省人民医院胃肠外科,四川成都610072 [2]成都职业技术学院医护分院护理教研室,四川成都610041

出　　处：《临床和实验医学杂志》2018年第13期1360-1363,共4页Journal of Clinical and Experimental Medicine

基　　金：四川省卫计委科研立项课题(编号:17PJ109)

摘　　要：目的探讨干扰p66shc基因表达对结直肠癌HCT116细胞增殖、迁移、迁移的影响。方法实验分为对照组(不做任何处理)、NC-siRNA组和p66SHC-siRNA组,NC-siRNA组、p66SHC-siRNA组通过Lipofectamine 2000将特异性siRNA转染入结直肠癌HCT116细胞中。荧光定量PCR(RT-PCR)和免疫印迹试验(Western Blot)检测细胞中p66SHC的表达量;四甲基偶氮唑(MTT)法检测细胞的增殖能力;划痕实验和Transwell法观察细胞迁移、侵袭的变化;Western Blot检测细胞EMT相关蛋白钙黏蛋白E(E-cadherin)、波形蛋白(Vimentin)、钙黏蛋白N(N-cadherin)的水平。结果采用siRNA干扰HCT116细胞中p66SHC蛋白的表达后,细胞的增殖能力显著下降(P<0.05),明显抑制细胞侵袭、迁移能力(P<0.05),E-cadherin蛋白的水平明显上调(P<0.05),Vimentin、N-cadherin蛋白的表达水平显著下调(P<0.05)。结论干扰p66SHC基因的表达能阻碍EMT的发生,从而降低结直肠癌细胞的增殖、侵袭以及迁移能力。Objective To investigate the effect of interfering with p66shc gene expression on the proliferation,migration and migration of colorectal cancer HCT116 cells. Methods The experiment were divided into control group,NC-siRNA group and p66SHC-siRNA group.Specific siRNA was transfected into colorectal cancer HCT116 cells by Lipofectamine 2000,and the expression of p66shc was detected by RT-PCR and Western Blot. The proliferation of cellswere detected by MTT assay. The migration and invasionwere observed by Scratch assay and Transwell method. Western Blot was used to detect the expression of E-cadherin,Vimentin,and N-cadherin. Results The siRNA had the significantly effects on inhibiting the expression of p66SHC in HCT116 cells（ P〈0. 05）,inhibited the invasion and migration of HCT116 cells（ P〈0. 05）,increased the protein levels of E-cadherin（ P〈0. 05）,decreased the protein levels of Vimentin and N-cadherin（ P〈0. 05）. Conclusion Interference of p66SHC gene expression can reverse the occurrence of EMT,and reduce the proliferation,invasion and migration of colorectal cancer cells.

关 键 词：结直肠癌 p66SHC基因 HCT116细胞 上皮细胞间充质化 

分 类 号：R735.34[医药卫生—肿瘤]