重庆医学Issue(32)：4288-4290,3.DOI:10.3969/j.issn.1671-8348.2014.32.005
MIA2慢病毒表达载体构建及其表达活性的研究
Construction of pLVX-IRES-ZsGreen1-MIA2 lentiviral expression vector and its expression activity
摘要
Abstract
Objective To construct an Lentiviral expression vector of pLVX‐IRES‐ZsGreen1‐MIA2 targeting to MIA2 and in‐vestigate its effect on the expression of MIA2 and growth of HCC cell line HepG2 in vitro ,observe MIA2 changes and the influence on apotheosis ,thus to provide preliminary experimental fundament for successive researching on the role of MIA2 in the pathogene‐sis of HCC .Methods The sequence of pLVX‐IRES‐ZsGreen1‐MIA2 was designed and synthesized .The pLVX‐IRES‐ZsGreen1‐MIA2 Lentiviral expression vector was constructed and then transiently transfected into HepG2 HCC cells in vitro .The proportion of pLVX‐IRES‐ZsGreen1‐MIA2 positive cells was observed under the fluorescence microscope .Then ,the expression level of MIA2 was detected by real time PCR .Moreover ,the proliferation of HepG2 cells was observed by MTT assay and colony formation as‐say .Finally ,the migration of HepG2 cells in vitro was also determined by Scratch assay .Results pLVX‐IRES‐ZsGreen1‐MIA2 Lentiviral expression vector was successfully constructed .Compared with control group (NC) ,the expression level of MIA2 was significantly decreased in transfected groups(P<0 .05);MTT assay showed that the proliferation of HepG2 cells was dramatically reduced in pIRES2‐ZsGreen1‐MIA2transfected groups(P< 0 .05);furthermore ,the number of both colony forming and migrating cells were also remarkably reduced in transfected groups(P<0 .05) .Conclusion The pIRES2‐ZsGreen1‐MIA2 can significantly re‐duce the expression level of MIA2 and inhibit the proliferation and migration of the HepG2 HCC cells in vitro .关键词
肝肿瘤/细胞增殖/黑色素瘤抑制蛋白2/HepG2细胞Key words
liver neoplasms/cell prdiferation/meanoma inhibitory activity related gene 2/HepG2 cells分类
医药卫生引用本文复制引用
杨花,杨少奇,何芳,胡建国,李鹏..MIA2慢病毒表达载体构建及其表达活性的研究[J].重庆医学,2014,(32):4288-4290,3.基金项目
国家自然科学基金资助项目（81060194）。 （）
