膜联蛋白A2对人肝癌细胞生物学行为的影响  

Effects of ANXA2 on Biological Behaviour of Human Hepotacellular Carcinoma in Vitro

作　　者：魏丽丽[1] 李润册[1] 强荣兵[1] 张楠楠[1] 刘月月[1] 张国芳[1] 杨梅[1] 侯颖春[1] 

机构地区：[1]陕西师范大学生命科学学院肿瘤分子细胞生物学实验室,西安710062

出　　处：《中国生物工程杂志》2010年第11期11-16,共6页China Biotechnology

基　　金：陕西省自然科学基础研究计划(SJ08-ZT09)资助项目

摘　　要：为了研究膜联蛋白A2(ANXA2)基因表达水平与人肝癌细胞生物学行为之间的关系,采用已优化转染条件的磷酸钙法将针对ANXA2的siRNA重组质粒导入人肝癌细胞SMMC-7721并观察对靶基因表达及细胞生物学行为的影响。以半定量RT-PCR和Western blotting检测转染后不同时间(24h、48h、72h和96h)ANXA2mRNA及蛋白表达变化;以MTT法、Hoechast33258染色及体外损伤修复实验等分析抑制ANXA2表达后对SMMC-7721细胞生物学行为的影响。结果显示:所设计的四条siRNA序列均不同程度抑制了ANXA2的表达(p<0.05),且呈时间依赖性;细胞生长能力及运动能力被显著抑制(p<0.05),其抑制效应与ANXA2表达敲低程度呈正相关。结论:ANXA2的高表达水平与肝癌细胞的凋亡、生长及运动能力密切相关,以RNAi方法抑制该基因的表达可以有效地抑制肝癌细胞的恶性生物学行为。上述结果为肝癌的实验性基因治疗研究提供了新的思路和对策。Objective：Investigate the correlation between ANXA2 and Hepatocellular Carcinoma（HCC）proliferation,apoptosis and motility.Method：Four siRNAs recombinants targeting to ANXA2 were introduced into SMMC-7721 cells using an optimized calcium phosphate transfection method.ANXA2 mRNA and protein levels were evaluated by using semi-quantitative RT-PCR and Western blotting.The cell proliferation,motility,and apoptosis were determined with MTT,wound healing assay,and Hoechast33258 staining methods.Results：Four siRNAs suppressed ANXA2 expression effectively with time dependence（p0.05）.Cell proliferation and motility were inhibited after ANXA2 knockdown（p0.05）,and the inhibitory efficiency was positively correlated with ANXA2 knockdown level.Conclusion：It is concluded that the over-expression of ANXA2 in HCC is closely associated with the cell malignant behaviors,and siRNA can inhibit the ANXA2 expression efficiently in SMMC-7721 cells.

关 键 词：膜联蛋白A2 RNA干扰 肝癌 生长、凋亡及迁移 

分 类 号：Q291[生物学—细胞生物学]