siRNA抑制cortactin基因表达对Hep-2细胞增殖和侵袭的影响  

Effects of silencing cortactin expression by siRNA on the proliferation and invasion in Hep-2 cells

作　　者：谭桂煌[1] 曾龙武[1] 侯素平[1] 张庆丽[1] 谢海龙[1] 

机构地区：[1]南华大学肿瘤研究所湖南,衡阳421001

出　　处：《中国癌症杂志》2010年第5期332-337,共6页China Oncology

摘　　要：背景与目的:皮层肌动蛋白(cortactin)是一种微丝肌动蛋白结合蛋白,其主要参与细胞骨架系统的调控,细胞外信号转导以及细胞黏附等过程,研究表明cortactin与肿瘤的侵袭和转移有关。因此,本研究旨在探讨siRNA(small interfering RNA)抑制cortactin基因表达对喉癌Hep-2细胞增殖和侵袭的影响。方法:构建含有cortactin基因的siRNA表达载体,转染Hep-2细胞,同时以转染空载体和空白细胞组作为对照(3组细胞分别命名为pSilencer3.1-cortactin-siRNA/Hep-2、pSilencer3.1/Hep-2和Hep-2),G418筛选稳定转染的细胞;Western blot检测siRNA对cortactin蛋白表达的影响;MTT和平皿克隆形成实验检测细胞增殖能力;Transwell法检测细胞迁移侵袭能力。结果:成功构建获得cortactin缺陷型稳定转染细胞系pSilencer3.1-cortactin-siRNA/Hep-2;与Hep-2细胞比较,pSilencer3.1-cortactin-siRNA/Hep-2组细胞中cortactin含量为11.22%(P<0.01),增殖被抑制,克隆形成率降低为21.47%(P<0.01),迁移、侵袭能力显著降低(P<0.01)。结论:cortactin表达下调能抑制喉癌Hep-2细胞的增殖和侵袭。Background and purpose：The cortical actin-binding protein,cortactin,participates in several functions in the cytoskeleton system,cellular signal transduction and cell adhesion.There is also increasing evidence that it regulates tumor invasion and metastasis.However,the role played by cortactin in laryngeal carcinoma has not been clearly delineated.The purpose of this experiment was to investigate the effect of silencing cortactin expression on the proliferation and invasion in the human laryngeal carcinoma cell line Hep-2.Methods：A plasmid from a siRNA targeting cortactin was constructed and transfected into a Hep-2 cell line.The siRNA interference efficiency of cortactin was determined by Western blot.The proliferation was measured by MTT assay and plate colony formation. The Transwell test was used to detect the migration and invasion ability of the Hep-2 cells.Empty plasmid-transfected Hep-2 and normal Hep-2 were used as control groups.Results：Compared to Hep-2 cells,the cortactin expression of pSilencer3.1-cortactin-siRNA/Hep-2 was 11.22%（P0.01）.Its proliferation was inhibited and its cloning efficiency was decreased to 21.47%（P0.01）.Its migratory and invasive ability was also significantly decreased（P0.01）. Conclusion：The expression of cortactin decreased could inhibit the proliferation and invasive ability of Hep-2 cells.

关 键 词：HEP-2细胞 SIRNA CORTACTIN 增殖 侵袭 

分 类 号：R730.261[医药卫生—肿瘤]