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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

23卷

期数:

2017年06期

页码:

510-515

栏目:

基础医学

出版日期:

2017-11-20

文章信息/Info

Title:

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

文章编号:

1006-8147(2017)06-0510-06

作者:

王多伟¹; Bikash Rai¹; 2; 王金淼¹; 戚 峰¹; 刘 彤¹

（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）

Author(s):

WANG Duo-wei¹;  Bikash Rai¹; 2;  WANG Jin-miao¹;  QI Feng¹;  LIU Tong¹

( 1. Department of General Surgery, General Hospital, Tianjin Medical University, Tianjin 300052, China; 2. Department of Surgery, Beijing United Family Hospitol, Beijing 100015, China)

关键词:

twist基因; 上皮间质转化; 结肠癌; 质粒; shRNA; 侵袭和转移

Keywords:

Twist gene;  epithelial-mesenchymal transition ;  colon carcinoma;  plasmid;  shRNA;  invasion and metastasis

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分类号:

R735.3+5

DOI:

-

文献标志码:

A

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pTracer-CMV/BSD-Twist 和pGenesil1.2-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。

Abstract:

Objective ：To explore the role of Twist gene in Epithelial-MesenchymalTransition (EMT) in SW480, HCT116 and HT29, and then study effect of Twist on invasion and metastasis of malignant tumors. Methods: Recombinant plasmids pTracer-CMV/BSD-Twist and pGenesil1.2-Twist-shRNA were used to transfect SW480, HCT116 and HT29. Transfection efficacy of the plasmid in each cell line was confirmed by flowcytometry. The mRNA 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 mRNA 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 mRNA 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). Conclusions: Up-regulation of Twist gene expression can promote the EMT and inhibiting the Twist gene expression can lessen the migration and invasion of HCT116.

参考文献/References:

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备注/Memo

备注/Memo:

基金项目天津医科大学总医院孵育基金项目（2YYFY2016021）

作者简介王多伟（1990-），男，硕士在读，研究方向：普通外科；

通信作者：戚峰， E-mail：qf@medmail.com.cn。

常用功能

更新日期/Last Update: 2017-11-14