慢病毒介导RNA干扰Pin1异构酶的鼻咽癌稳定细胞株的建立和鉴定  

Identification of stable expression cells of nasopharyngeal carcinoma by a lentivirus for RNA interference( RNAi) of Pin1

作　　者：陈华[1] 黄国良[1] 何志巍[1] 

机构地区：[1]广东医学院中美肿瘤研究所及广东省医学分子诊断重点实验室,广东东莞523808

出　　处：《现代肿瘤医学》2015年第7期889-892,共4页Journal of Modern Oncology

基　　金：国家自然基金面上项目(编号:81372137)

摘　　要：目的:应用慢病毒介导的RNA干扰技术,建立稳定表达si Pin1的鼻咽癌细胞株。方法:将对照组病毒液lv-3和实验组病毒液si Pin1感染CNE2细胞,通过嘌呤霉素和绿色荧光检测筛选si Pin1稳定细胞株,在荧光显微镜下观察细胞的荧光表达,通过定量PCR和Western blot鉴定si Pin1细胞Pin1 mRNA和蛋白的表达。结果:筛选si Pin1稳定表达细胞株嘌呤霉素筛选浓度为1μg/ml,病毒感染后超过90%细胞发出绿色荧光,实验组si Pin1与对照组lv-3相比,Pin1 mRNA表达水平下降,Western blotting检测Pin1蛋白表达明显减少。结论:成功构建Pin1干扰的稳定表达鼻咽癌细胞株,为后续研究提供工具细胞。Objective： To identify a stable expression cell of nasopharyngeal carcinoma by a lentivirus for RNA interference（ RNAi） of the Pin1. Methods： The lentivirus packaging mix lv- 3 and si Pin1 were transfected into CNE2 cells and inspected by fluorescence microscopy. Puromycin was used to screen positive cells. The expression of Pin1 mRNA and protein was detected by RT- PCR and Western blotting respectively. Results： The lentivirus lv- 3 and si Pin1 were screened with 1μg / ml puromycin. The expression of Pin1 mRNA and protein was downregulated. Conclusion： A stable expression cell of nasopharyngeal carcinoma was successfully constructed and could be used as a tool for the following studies.

关 键 词：PIN1 RNA干扰 慢病毒 鼻咽癌 

分 类 号：R730[医药卫生—肿瘤] R739.63[医药卫生—临床医学]