文献类型：期刊文献

中文题名：凋亡素融合蛋白在HeLa细胞中的转导及体外活性分析

英文题名：Apoptin Fusion Protein Transduced in HeLa Cells and Its Activity Assay in vitro

作者：华迪[1];赵健[1];范立强[1];王富军[2];汪雅雯[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237;[2]浙江日升昌药业有限公司,浙江东阳322100

年份：2010

卷号：36

期号：5

起止页码：651

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;Scopus;北大核心:【北大核心2008】；CSCD:【CSCD2011_2012】；

语种：中文

中文关键词：穿膜肽;EC-SOD;HeLa细胞

外文关键词：cell-penetrating peptides; EC-SOD; HeLa cell

摘要：研究胞外超氧化物歧化酶(EC-SOD)的肝素结合域(HBD)在HeLa细胞中的转导活性,并证实HBD具有运载药物蛋白(凋亡素)进入细胞并诱导肿瘤细胞凋亡的能力。通过克隆表达的HBD-EGFP融合蛋白经Ni2+-NTA纯化后,观察其转导活性;并利用HBD-凋亡素融合蛋白表达纯化后,用噻唑蓝(MTT法)检测肿瘤细胞的凋亡。结果表明:用荧光显微镜可观察到HeLa细胞内有绿色荧光;MTT法检测表明凋亡素融合蛋白能诱导HeLa细胞的凋亡;HBD具有转导活性,能携带药物蛋白(凋亡素)进入细胞并能诱导HeLa细胞的凋亡。
To study transduction activity of the heparin-binding domain（HBD）of extracellular superoxide dismutase（EC-SOD）in HeLa cell,and to confirm HBD have the ability to deliver pharmaceutical proteins（apoptin） into cells and to induce apoptosis of tumor cells.Cloned HBD-EGFP fusion protein was expressed and purified by Ni^（2＋）-NTA affinity chromatography,to observe its transduction activity.HBDapoptin fusion protein was expressed and purified,then the apoptosis of tumor cells in vitro was tested by MTT assay.The results show that green fluorescence can be observed in HeLa cells using fluorescence microscopy.MTT results indicated that fusion protein of apoptin can induce apoptosis of HeLa cells.HBD with the transduction activity,was capable of carrying drugs protein（apoptin） to enter cells and can induce apoptosis in HeLa cells.