Twist基因对结肠癌细胞系侵袭转移能力影响的体外研究  

Research on the influence of Twist gene on invasion and metastasis of colon cancer cell lines in vitro

作　　者：王多伟 Bikash Rai 王金淼[1] 戚峰[1] 刘彤[1] 

机构地区：[1]天津医科大学总医院普通外科,天津300052 [2]北京和睦家医院外科,北京100015

出　　处：《天津医科大学学报》2017年第6期510-515,共6页Journal of Tianjin Medical University

基　　金：天津医科大学总医院孵育基金项目(2YYFY2016021)

摘　　要：目的:探讨Twist基因在SW480、HCT116和HT29结肠癌细胞系上皮-间质转化(EMT)中的作用,明确其对恶性肿瘤侵袭转移能力的影响。方法:利用重组质粒p Tracer-CMV/BSD-Twist和p Genesil1.2-Twist-shRNA转染SW480、HT29和HCT116;流式细胞术检测转染率;Real time-PCR和Western blot检测Twist、E-cadherin和Vimentin转录及蛋白表达水平;Transwell实验检测细胞的迁移和侵袭能力。结果:转染高表达Twist质粒后,各结肠癌细胞系中Twist和Vimentin表达水平显著升高(P<0.05),E-cadherin显著降低(P<0.05);转染低表达Twist质粒后,SW480和HT29中各指标均无显著变化(P>0.05),HCT116中Twist和Vimentin表达水平显著降低(P<0.05),E-cadherin无显著变化(P>0.05);Transwell实验表明抑制HCT116细胞系Twist表达后,其侵袭和迁移能力明显减弱(P<0.01)。结论:上调Twist表达可以促进EMT;抑制HCT116中Twist表达能减弱结肠癌细胞的侵袭和转移能力。Objective： To explore the role of Twist gene in epithelial-mesenchymal transition（EMT） in SW480, HCT116 and HT29, and then study the effect of Twist on invasion and metastasis of malignant tumors. Methods： Recombinant plasmids p Tracer-CMV/BSD-Twist and p Genesil1.2-Twist-shRNA were used to transfect SW480, HCT116 and HT29. Transfection efficacy of the plasmid in each cell line was confirmed by flowcytometry. The m RNA transcription level and protein expression level of Twist, E-cadherin and Vimentin were detected by RT-PCR and western blot, respectively. The migration and invasive analysis was done by transwell assay. Results： After transfected by recombinant high-expressed twist plasmid, the m RNA and protein expression levels of Twist and Vimentin increased significantly（P〈0.05）, whereas E-cadherin was inhibited（P〈0.05）. After transfected by recombinant low-expressed twist plasmid, all the parameters in SW480 and HT29 cell lines were statistically insignificant（P〉0.05）, the m RNA and protein levels of Twist and Vimentin were prominently inhibited in HCT116 cell line（P〈0.05） and the level of E-cadherin was not statistically significant（P〈0.05）. Significant reduction of invasion and migration was observed in transwell assay after inhibiting the Twist expression in HCT116 cell line（P〈0.01）.Conclusion： Up-regulation of Twist gene expression can promote the EMT and inhibiting the Twist gene expression can lessen the migration and invasion of HCT116.

关 键 词：TWIST基因 上皮间质转化 结肠癌 质粒 SHRNA 侵袭和转移 

分 类 号：R735.35[医药卫生—肿瘤]