shRNA对肺癌相关基因抑制作用的实验研究  

The experimental study of short hairpin RNA on the suppression of lung cancer-associated gene

作　　者：徐鹏飞[1] 王秦秦[2] 朱中山[3] 

机构地区：[1]陕西中医学院,陕西咸阳712000 [2]昆明医学院第一附属医院,云南昆明650032 [3]湖南省第二人民医院肿瘤科,湖南长沙410006

出　　处：《现代肿瘤医学》2007年第8期1071-1073,共3页Journal of Modern Oncology

基　　金：国家十五科技攻关项目(编号2001BA703B11)

摘　　要：目的:利用RNA干扰(RNAi)技术,研究突变型蛋白磷酸酶2A(PP2A)α催化亚基编码基因的小发夹RNA(shRNA)对人肺癌GLC-82细胞的影响。方法:根据靶基因的不同区域设计4组shRNA,采用体外转录法合成,利用阳离子脂质体转染入GLC-82细胞,孵育48小时后,观察细胞形态学改变,MTT法检测细胞的存活率,免疫印迹法检测蛋白质的表达。结果:与空白对照相比,4组shRNA作用不尽相同,其中一组shR-NA可以明显抑制细胞的增殖,出现部分细胞脱落死亡,并且抑制GLC-82细胞目的蛋白的表达。结论:成功筛选出一段能特异而高效地抑制突变型PP2Aα催化亚基基因表达的shRNA,为进一步研究该基因的功能和作用机制提供了新的手段。另外,应用RNA干扰技术可抑制突变型PP2Aα催化亚基编码基因在人肺癌GLC-82细胞中的表达,有望成为肺癌基因治疗新的研究线索。Objective：To study the effect of encoding gene of short hairpin RNA （shRNA） targeting alpha catalytic submit of mutant protein phosphatase 2A （PP2A） on GLC -82 cells with RNA interference technology. Methods： Four shRNA targeting the mRNA of mutant protein phosphatase 2A were synthesized by in vitro transcripition and transfected into GLC - 82 cells with lipufection. After incubation for 48 hours, we observed and evaluated the cells morphological change , the survival rate of the cells with MTT assay and the expression with Western blot. Results： Compared with the blank control,the four shRNA had not the same effects, One shRNA suppressed cell prolifarition and some cells dead, and showed obvious effectes on inhibiting the protein expression of GLC - 82 cells. Conclusion： One shRNA of the four were obtained, which could specially and effectively inhibit the expression of alpha catalytic submit of mutant protein phosphatase 2A in GLC - 82 cells. It may open a new approach to use shRNA as a new tool to study this gene＇s function and mechanisms, and may be develloped to be a new gene therepautic agent for lung cancer.

关 键 词：SHRNA RNAI 突变型 磷酸酶2Aα催化亚基 

分 类 号：R734.2[医药卫生—肿瘤]