MDA-7/IL-24真核表达载体的构建和重组腺病毒的制备及其对肝癌细胞生长抑制作用的研究  

Construction and expression of recombinant adenovirus encoding MDA-7/IL-24 and its inhibitory effect on proliferation in human hepatocellular carcinoma cells

作　　者：张翔[1,2] 孟艳玲[1,3] 王磊[1,2] 赵晶[1,2] 闫博[1,3] 陈锐[1,2] 张瑞[1,2] 杨安钢[1,2,3] 

机构地区：[1]第四军医大学肿瘤生物学国家重点实验室,陕西西安710032 [2]第四军医大学生物化学与分子生物学教研室,陕西西安710032 [3]第四军医大学免疫学教研室,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2011年第12期1277-1279,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(81172149;30873006;30901771)

摘　　要：目的:构建含有MDA-7基因的的真核表达载体并制备重组腺病毒,转染/感染肝癌细胞后,观察其对细胞增殖的影响。方法:构建MDA-7的真核表达载体,将表达质粒转染肝癌细胞,利用平板克隆形成实验观察MDA-7分子对肿瘤细胞系克隆形成能力的影响。利用Adeasy-1腺病毒重组系统构建、包装并扩增含有MDA-7基因的重组腺病毒。利用MTT实验检测Ad-MDA-7对肿瘤细胞增殖的影响。结果:成功构建了MDA-7真核表达载体,利用平板克隆形成实验证实其可抑制肝癌细胞的克隆形成能力。成功地构建并包装了含有MDA-7基因的重组腺病毒,利用MTT实验证明其可抑制肝癌细胞的增殖。结论:MDA-7对肝癌细胞的增殖具有显著的抑制作用,为进一步研究其在肝癌细胞中的功能提供了重要的实验依据。AIM： To construct recombinant plasmid and adenovirus harboring MDA-7 gene, and to investigate its biological function on the proliferation of human hepatocelJular carcinoma cells. METHODS： The MDA-7 fragments from the T vectors were inserted into pCDNA-3 vector to construct expression plasmids named pCDNA3-MDA-7. To determine its effects on the proliferation of HCC cells, transfected the expression vector into cells and tested the ability of colony formation in cancer cells. Simultaneously, con- structed recombinant adenovirus expressing MDA-7, and detected its effect on the proliferation of HCC cells by using MTT- assay. RESULTS, Successfully constructed plasmid- and adenovirus-based system to express MDA-7. The data of colony formation assay and MTT- test showed that MDA-7 can obviously suppress cell growth in HCC cells. CONCLUSION： MDA- 7 may function as tumor suppressor in HCC cells, and the adenovirus-mediated MDA-7 can be a novel strategy for the anti- HCC therapy. Our study established the foundation for future research on the effect of MDA-7 in HCC.

关 键 词：黑色素瘤分化相关抗原-7 肝癌 腺病毒 细胞增殖 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]