Twist促进人肝癌细胞的迁移  

Twist Promotes Migration of Hepatocellular Carcinoma Cells

作　　者：杜艳涛 韩海勃[1] 赵威[1] 张志谦[1] 

机构地区：[1]北京大学临床肿瘤学院,北京肿瘤医院暨北京市肿瘤防治研究所临床实验室,恶性肿瘤发病机制及转化研究教育部重点实验室,北京市100142

出　　处：《中国肿瘤临床》2010年第7期361-364,共4页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金(编号:30671060,30971494);教育部新世纪优秀人才支持计划项目资助(编号:NCET-07-0031)~~

摘　　要：目的:研究Twist对肝癌细胞迁移能力的影响。方法:RT-PCR方法检测Hep-11和Hep-12中Twist的表达情况。将Twist-cDNA转染Hep-11筛选得到稳定高表达Twist的细胞克隆Twist-Hep-11。构建pSuper-Twist-shRNA干扰载体,瞬时转染Hep-12细胞。用Boyden小室检测过表达和抑制Twist基因后对细胞迁移和侵袭能力的影响。RT-PCR和Western blot检测Hep-11上调Twist后的EMT标志分子的表达。结果:与Hep-11相比,Hep-12的Twist基因表达显著上调,Hep-12的迁移能力也强于Hep-11。Hep-11细胞过表达Twist后,其迁移能力明显增强;而干扰Hep-12细胞Twist表达后,其迁移能力亦明显下降。Hep-11过表达Twist后,N-cad、Vimentin表达明显上调,E-cad变化不明显。结论:Twist可以促进肝癌细胞的迁移,其机制可能与N-cad、Vimentin表达上调有关。Objective： To study the effect of Twist gene on migration of human hepatocellular carcinoma cells and the possible mechanisms involved. Methods： RT-PCR was used to detect expression of Twist gene in Hep11 and Hep12 cell lines. Hep11 cells were stably transfected with Twist-cDNA, and Hep12 cells were transiently transfected with Twist RNAi plasmid. Cell migration assay was performed on Twist upregulated Hep11 cells and Twist RNAi Hep12 cells. RT-PCR and Western blot were performed to detect the expression of epithelial mesenchymal transition （EMT） markers. Results： Compared with that in primary Hep11 cells, Twist was highly expressed in recurrent Hep12 cells. Cell models （Twist-Hep11） with steady over-expression of Twist protein were obtained. Compared with pcDNA3-Hep11 cells, the migration of Twist-Hep11 cells were obviously increased. However, the migration of Twist RNAi Hep12 （Si-Twist-Hep12） cells was reduced. Overexpression of Twist in Hep11 cells promoted N-cadherin and Vimentin expression. Conclusion： Twist promotes the migration of hepatocellular carcinoma cells in vitro and may play an important role in upregulation of mesenchymal markers in hepatocellular carcinoma.

关 键 词：TWIST RNA干扰 基因转染 迁移 侵袭 

分 类 号：R977.21[医药卫生—药品]