南京医科大学学报（自然科学版）2011，Vol.31Issue(2)：161-165,5.
人膜连蛋白A2基因重组慢病毒载体的构建及其在肝癌细胞株中的表达
Construction of the recombinant lentiviral vector containing human ANXA2 gene and its expression in hepatocellular carcinoma cells
摘要
Abstract
Objective: To construct the recombinant lentiviral vector containing human annexin A2(ANXA2) gene and measure the expression level of ANXA2 in hepatocellular cell line of Hepa 1-6. Methods:The ANXA2 fragment was amplified by PCR and subcloned into the lentiviral vector pWPT. The resultant lentivirus were confirmed by PCR, restriction enzyme digestion and DNA sequencing. Recombinant lentivirus (LV-ANXA2-Flag) were produced from 293T by transient calcium-phosphate co-transfection with pWPF-ANXA2-Flag, CMVRS.91and pCMV-VSV-G. Hepa 1-6 cells were infected by LV-ANXA2-Flag lentivirus, and the expression of ANXA2 was confirmed by RT-PCR and Western blotting. Cell cycles were measured via flow cytometry. Results:The recombinant lentiviral vector carried the ANXA2 gene was successfully constructed. RT-PCR and Western blot analysis revealed that the ANXA2 gene can be correct transcripted and translated in Hepa 1-6 cell which infected by LV-ANXA2-Flag. After infected,the Hepa 1-6 cells were higer than control cells in percentage of S phase cells. Conclusion: The recombinant lentiviral vector pWPT-ANXA2-Flag was constructed successfully, it could be used to transfect hepatocellular carcinoma cells. ANXA2 could be stablely expressed and promote the proliferation of Hepa 1-6 cell.关键词
膜连蛋白A2/慢病毒载体/肝癌Key words
annexin A2/ lentiviral vector/ hepatocellular carcinoma/分类
医药卫生引用本文复制引用
杨东昌,母小新,刘巧玉,邓蕾,陈云,孙倍成..人膜连蛋白A2基因重组慢病毒载体的构建及其在肝癌细胞株中的表达[J].南京医科大学学报（自然科学版）,2011,31(2):161-165,5.基金项目
国家自然科学基金青年基金(30901750) （30901750）
南京医科大学科技发展基金重点项目(NMUZ009) （NMUZ009）
教育部"新世纪优秀人才支持计划"(NCET-09-0160) （NCET-09-0160）
江苏省卫生厅"科教兴卫工程"医学重点学科(RC2007057) （RC2007057）
