靶向敲低核蛋白1抑制HepG2肝癌细胞的增殖及迁移  

Nuclear protein 1 knockdown inhibits proliferation and migration of Hep G2 cells

作　　者：黄红艳[1] 张彦斌[1] 王小利[1] 周心娜[1] 李莎[1] 王嫚娜 任军[1] 

机构地区：[1]首都医科大学附属北京世纪坛医院肿瘤内科,肿瘤治疗性疫苗北京市重点实验室,北京100038 [2]军事医学科学院生物工程研究所,北京100071

出　　处：《细胞与分子免疫学杂志》2015年第6期782-786,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金面上项目(81172534,81472588)

摘　　要：目的观察应激相关核蛋白1(Nupr1)在不同肝癌细胞系中的表达,通过靶向敲低Hep G2肝癌细胞Nupr1,观察其对肝癌细胞增殖及迁移的影响。方法通过实时定量PCR检测BEL-7402、QSG-7703、SMMC-7721、Hep G2肝癌细胞中Nupr1的表达。采用RNA干扰技术敲低Hep G2细胞中Nupr1的表达,通过MTT法及集落形成实验比较细胞的增殖能力的变化,用流式细胞术进行细胞周期分析,利用TranswellTM实验观察Nupr1对细胞迁移能力的影响。结果 Hep G2人肝癌细胞中Nupr1的表达显著高于其他肝癌细胞系。靶向Nupr1的2个短发夹RNA(shRNA)均能有效敲低Hep G2肝癌细胞中Nupr1的表达,抑制效率分别为72.25%和84.25%。Nupr1表达降低能够抑制细胞增殖,导致细胞周期发生G1期阻滞。Nupr1敲低显著降低细胞的迁移能力及集落形成能力。Western blot结果显示Nupr-1表达降低能够上调细胞周期负性调控因子p21和p27的表达水平。结论靶向敲低Nupr1抑制Hep G2肝癌细胞的增殖及迁移。Objective To observe the expression of nuclear protein 1 （Nuprl）, a stress-related nuclear protein, in a panel of human hepatocellular carcinoma cell lines, and its effects on the proliferation and migration of hepatocellular carcinoma cells by RNA interference-mediated knockdown. Methods Real-time quantitative PCR was employed to detect Nuprl mRNA levels in hepatocellular carcinoma cell lines, including BEL-7402, QSG-7703, SMMC-7721 and HepG2. After Nuprl expression was knocked down by RNA interference, cell proliferation was monitored by M3-r assay and colony formation in plate. Cell cycle was analyzed by flow cytometry and cell migration was assayed by TranswellTM assay. Resells Higher expression of Nuprl was detected in HepG2 as compared with the other cell lines. Nuprl expression in HepG2 cells were efficiently knocked down by two short hairpin RNAs （shRNAs）, with inhibitory rates being 72.25% and 84.25%, respectively. HepG2 cells with Nuprl knockdown displayed lower rate of proliferation, GI arrest, and significantly decreased abilities of cell migration and colony formation. Western blotting showed that Nuprl knockdown increased the expressions of io21 and p27, two negative regulators of cell cycle. Conclusion Knockdown of Nuprl inhibited the proliferation and migration of HepG2 hepatocellular carcinoma cells.

关 键 词：肝癌 核蛋白1 增殖 迁移 细胞周期 

分 类 号：R735.7[医药卫生—肿瘤] Q255[医药卫生—临床医学]