摘要
目的:利用抗死亡受体5(DeathReceptor5)单克隆抗体(mAb),检测DR5在肝癌细胞系的表达。方法:sDR5免疫BALB/c小鼠,小鼠脾细胞与SP2/0-Ag14细胞在50%PEG条件下融合,获得能够分泌抗DR5单克隆抗体的杂交瘤细胞株,利用杂交瘤细胞株分泌的抗DR5特异性单克隆抗体,采用流式细胞仪技术测定抗DR5mAb结合至细胞表面的量来分析死亡受体DR5在肝癌细胞系及肝细胞表面的表达情况。结果:不同细胞表面DR5的表达水平分别为:人肝癌SMMC7721细胞95%、人肝细胞癌HepG2细胞40%,人正常肝细胞HL-770220.3%。结论:抗DR5mAb能够特异性与DR5结合。DR5在人肝癌SMMC7721细胞高水平表达,人肝细胞癌HepG2细胞中度表达,人正常肝细胞HL-7702细胞低表达。该特异性mAb在研究TRAIL受体的生物功能上是一种有用的工具,对研究DR5在组织和细胞系表达十分有价值。
Objective： To establish hybridomas that produces anti-death receptor 5 mAbs and checks surface expression of death receptor on cell lines. Methods： BALB/c mice were immunized with sDR5 in CFA. The spleen cells were fused with the myeloma SP2/0-Ag 14 in the presence of 50% polyethylene glycol. Surface expression of TRAIL receptor was determined by flow cytometric analysis by measuring the binding of anti-DR5 mAb. Results： The expression level of the different cell surface DR5 were 95% on SMMC7721 cells, 40% on HepG2 cells and 20.3% on HL7702 cells. Conclusions： These results demonstrate that anti-DR5 mAb is able to specifically bind to DR5 and that DR5 is expressed at high level on SMMC7721 cell lines and middle level on HepG2 cell lines. The expression of DR5 on HL-7702cell lines is low. The DR5 specific mAb described here will serve as a valuable tool for studying the biology of TRAIL the receptors. The anti-DR5 mAb generated in the present study may be useful for investigating DR5 expression in tissues or cell lines.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2006年第5期287-289,共3页
Chinese Journal of Clinical Oncology
基金
河南省杰出人才创新基金(编号:96-906-01-23)
河南省医学科技创新人才工程基金项目资助(编号:96-906-01-23)
