腺病毒介导mda-7/IL-24基因感染对卵巢癌耐药细胞凋亡的影响  

Effect of Adenovirus-mediated mda-7/IL-24 Gene Infection on Apoptosis of Drug-resistant Human Ovarian Cancer Cell Lines OVCAR-3 and OVCAR-8/TR

作　　者：熊炬[1] 彭芝兰[1] 谭欣[1] 闫乃红[2] 

机构地区：[1]四川大学华西第二医院妇产科,四川成都610041 [2]四川大学华西医院眼科分子遗传实验室,四川成都610041

出　　处：《癌症》2007年第4期371-376,共6页Chinese Journal of Cancer

摘　　要：背景与目的:Mda-7/IL-24基因是一个具有选择性的诱导肿瘤细胞凋亡和细胞因子免疫调节功能双重作用的基因,在抗肿瘤基因治疗方面有很好的应用前景,本研究利用构建的mda-7/IL-24重组腺病毒感染卵巢癌耐药细胞株,了解其对卵巢癌耐药细胞的抗肿瘤作用。方法:利用腺病毒AdEasy1载体构建含目的基因的Ad.mda-7/IL-24,感染两种卵巢癌耐药细胞株OVCAR-3和OVCAR-8/TR,Westernblot法检测感染后MDA-7/IL-24蛋白表达,用Hoechst33258凋亡染色和流式细胞仪检测感染后细胞的凋亡和细胞周期改变。结果:Ad.mda-7/IL-24经测序、酶切电泳、感染后Westernblot检测MDA-7蛋白表达,表明构建成功;感染Ad.mda-7后72h内OVCAR-3细胞最高凋亡率为14.1%,OVCAR-8/TR细胞为32.4%,明显高于空载体组和未感染组。结论:成功构建了Ad.mda-7/IL-24重组腺病毒,用其感染卵巢癌耐药细胞可诱导细胞凋亡。BACKGROUND ＆ OBJECTIVE： Melanoma differentiation associated gene-7 （mda-7/IL-24） has double functions： specifically induces tumor cell apoptosis and modulates immune responses. Therefore, it is a strong candidate for human cancer gene therapy. This study was to evaluate the effect of adenovirus-mediated mda-7/IL-24 infection on the apoptosis of drug-resistant ovarian cancer cell lines OVCAR-3 and OVCAR-8/TR. METHODS： Adenovirus-mediated mda-7/IL-24 （Ad.mda-7/IL-24） was constructed using AdEasy 1 system. OVCAR-3 and OVCAR-8/TR cells were infected by Ad.mda-7/IL-24. The expression of MDA-7/IL-24 protein was detected by Western blot. Cell apoptosis was detected by flow cytometry with Hoechst33258 staining. Cell cycle distribution was detected by flow cytometry. RESULTS： The recombinant Ad.mda-7/IL-24 was confirmed by DNA sequencing and electrophoresis. The expression of MDA-7/IL-24 protein was detected in the cells after infection. Within 72 h after Ad.mda-7/IL-24 infection, the maximal apoptosis rates of OVCAR-3 and OVCAR-8/TR cells were 14.1% and 32.4%, respectively, significantly higher than empty vector group and uninfected group. CONCLUSIONS： The recombinant Ad.mda-7/ IL-24 was successfully constructed. It can induce apoptosis in drug-resistant ovarian cancer OVCAR-3 and OVCAR-8/TR cells.

关 键 词：腺病毒 感染 卵巢肿瘤 耐药细胞株 凋亡 

分 类 号：R737.31[医药卫生—肿瘤]