抗HER2/neu单链抗体增强TNF-α对卵巢癌细胞的抗肿瘤活性  

Anti-HER2/neu Augments TNF-α-induced Biological Effects in Ovarian Cancer Cell Line that Overexpresses HER2/neu

作　　者：蒋琳[1] 纪剑飞 阎锡蕴[3] 吕安国[1] 吴文芳[1] 

机构地区：[1]中国科学院沈阳应用生态研究所 [2]Beckman Research Institute, City of Hope National Medical Center, 1500 Duarte Rd. Duarte, CA 91010, USA [3]中国科学院生物物理研究所北京100101

出　　处：《应用与环境生物学报》2006年第4期543-546,共4页Chinese Journal of Applied and Environmental Biology

基　　金：沈阳市科委项目(No.20011221-03);中国科学院沈阳应用生态所与屹昌科技集团股份有限公司合作项目~~

摘　　要：为了探索抗HER2/neu单链抗体与TNF-α联合应用对表面过度表达HER2/neu卵巢癌细胞的生物效应,同时构建了抗HER2/neu单链抗体scFvC6.5的原核表达载体和人TNF-α的原核表达载体,将上述两种重组子分别转化入感受态宿主菌BL21(DE3),得到稳定表达.表达产物主要以包含体形式存在;包含体经过溶解、变性、复性和纯化,得到了分纯的产物.SDS-PAGE和Western-blot检测结果证实,蛋白表达正确.ELISA法验证了scFvC6.5和人TNF-α具有与卵巢癌细胞SKOV-3的结合活性.MTT细胞毒活性试验进一步表明,相对于单独使用TNF-α,联合应用上述两个重组蛋白细胞毒效应明显提高,SKOV-3细胞对TNF-α的敏感性增强.这将为抗HER2/neu抗体的联合抗肿瘤疗法提供一种新的途径,具有潜在的临床应用价值.It was investigated whether combined treatment with tumor necrosis factor-α and anti-HER2/neu scFvC6.5 could enhance the specific killing of ovarian cancer cell line that overexpresses HER2/neu. The scFvC6.5 or TNF-α gene fragment was cloned into the pET28a expression vector, and two recombinant plasmids were transformed into Escherichio coli BL21 （DE3） , respectively. Proteins were purified by Ni-NTA chromatography column. The SDS - PAGE and Western-blot assay confirmed the correct protein expression. Binding study of either scFvC6.5 or TNF-α to ovarian cancer cell line SKOV-3 was assessed by ELISA. The study on cytotoxieity against SKOV-3 cells demonstrated that the combined treatment with tumor necrosis faetor-α and scFvC6.5 was more active than free TNF-α. These data suggested that the combination of scFvC6.5 and TNF-α made the anti-tumor activity and specificity enhanced in the treatment of HER2/neu-overexpressing tumors.

关 键 词：抗HER2/neu单链抗体 肿瘤坏死因子 基因表达 联合应用 

分 类 号：R737.31[医药卫生—肿瘤] R730.5[医药卫生—临床医学]