EGFP-Apoptin融合蛋白诱导肿瘤细胞凋亡的研究  

Apoptosis of Tumor Cells Induced by Fusion Protein EGFP-Apoptin

作　　者：寇冰[1] 彭冬君[1] 孙军[1] 田俊[1] 宗义强[1] 屈伸[1] 

机构地区：[1]华中科技大学同济医学院基础医学院生物化学与分子生物学系,武汉430030

出　　处：《华中科技大学学报（医学版）》2006年第3期281-283,F0004,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省自然科学基金资助项目(No.2001ABA004)

摘　　要：目的 观察EGFP-Apoptin融合蛋白在肿瘤细胞中的分布和亚细胞定位，探讨Apoptin诱导肿瘤细胞凋亡的机制。方法 构建EGFP-Apoptin融合蛋白真核表达载体，用脂质体转染技术将其导入人肝癌细胞株HepG2中，在荧光显微镜下观察Apoptin在肿瘤细胞中的分布、亚细胞定位；用TUNEL法检测其在体外诱导肿瘤细胞凋亡的效应。结果 转染细胞中EGFP-Apoptin在肿瘤细胞中得以高表达，转染24h后逐渐从细胞质迁移至细胞核，最后定位于细胞核内。转染4～5d后逐渐诱导肿瘤细胞凋亡。结论 EGFP-Apoptin融合蛋白在肿瘤细胞中具有核定位效应，并诱导肿瘤细胞凋亡；EGFP-Apoptin可以作为研究Apoptin在肿瘤细胞中分布和亚细胞定位的有效工具，用以探讨Apoptin在体外诱导肿瘤细胞凋亡的机制。Objective To observe the distribution and subcellular location of fusion protein EGFP-Apoptin in tumor cells, and to study the mechanism of Apoptin-induced apoptosis of tumor cells. Methods The eukaryotic expression vector of EGFP- Apoptin was constructed and transfected into human bepatocarcinoma cell line HepG2 by liposome transfection. The distribution and subcellular location of fusion protein EGFP-Apoptin in tumor cells were observed under fluorescence microscopy. The EGFP-Apoptin induced apoptosis of tumor cells in vitro was tested by TUNEL assay. Results High expression of EGFP-Apoptin was detected in the transfected tumor cells, and EGFP-Apoptin moved from cytoplasm to nucleus gradually after 24 b, finally accumulated in nucleus. Apoptosis of tumor cells was induced on the 4th-5th day after transfection. Conclusion The fusion protein EGFP-Apoptin has nucleic localization function in tumor cells and can induce the apoptosis of tumor cells. EGFP- Apoptin can be used as an effective tool for the research of distribution and location of Apoptin to explore the mechanism of Apoptin inducing apoptosis of tumor cells in vitro.

关 键 词：凋亡素 细胞凋亡 增强型绿色荧光蛋白 

分 类 号：R730.23[医药卫生—肿瘤]