靶向磷脂酰肌醇蛋白聚糖3的小干扰RNA筛选和对肝癌细胞的作用  

Selection of small interfering RNA targeting Glypican-3 and its influence on liver cancer cells

作　　者：邱志东[1] 缪辉来[1] 雷长江[1] 刘忠考[1] 黎然[1] 包仕廷[1] 

机构地区：[1]广东医学院附属医院肝胆外科,广东湛江524001

出　　处：《中华实验外科杂志》2012年第9期1727-1729,共3页Chinese Journal of Experimental Surgery

基　　金：广东省科技计划资助项目（20101700501）;广东省医学科研基金资助项目（B2011242）

摘　　要：目的探讨磷脂酰肌醇蛋白聚糖3（GPC3）在肝细胞癌（HCC）发生发展中的作用机制。方法设计靶向GPC3的siRNA，转染Huh7细胞后实时荧光定量聚合酶链反应（Real-timePCR）检测其对GPC3表达的抑制效果；选用抑制效果明显的小干扰RNA（siRNA）下调GPC3表达后，Westernblot法检测GPC3的蛋白表达水平，并用流式细胞仪、荧光显微镜观察及Transwell法分别对Huh7细胞的周期、增殖和侵袭进行检测。结果成功筛选出1条抑制效率达到74．87％的特异靶向GPC3的siRNA，可以显著抑制GPC3的转录翻译；同时，可见Huh7细胞的增殖和侵袭受到抑制，但并不影响细胞周期。结论GPC3的表达有利于HCC的增殖侵袭，其表达降低不利于HCC的发生发展。Objective To study the action mechanism of Glypican-3 （GPC3） in the development of hepatocellular carcinoma （HCC）. Methods siRNA targeting GPC3 was designed and transfected into Huh7 ceils, and the inhibitory effects on the expression of GPC3 was tested by using real-time polymerase chain reaction （PCR）. The siRNA with obvious inhibitory effect was selected to down-regulate the GPC3 expression, and then the expression of GPC3 protein was detected by usig Wesnter blotting. The cell cycle, proliferation and invasion of Huh7 cells were assayed by using flow cytometry, fluorescence microscopy and Transwell, respectively. Results We successfully screened out a siRNA specifically targeting GPC3, its inhibition efficiency was 74. 87% and the siRNA could significantly inhibit the transcription and translation of GPC3. The knock-down of GPC3 could inhibit the proliferation and invasion of Hut17 cells, but had no effects on the cell cycle. Conclusion GPC3 expression facilitates the proliferation and invasion of HCC.

关 键 词：癌 肝细胞 磷脂酰肌醇蛋白聚糖3 小干扰RNA HUH7 

分 类 号：R735.7[医药卫生—肿瘤]