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.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
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)
作者简介
寇冰，男，1966年生，博士研究生
通讯作者，Corresponding author，Tel：027-83692624 E-mail：qushen@mails．tjmu．edu．cn
