sFv-TNF-α基因修饰的PBMCs对人肝癌细胞系HHCC的体外杀伤活性研究  

Enhanced in vitro cytotoxicity of PBMCs via genetic modification of single-chain bifunctional antibody

作　　者：程虹[1] 刘彦仿[1] 张惠中[2] 沈万安[2] 杨安钢[3] 马福成[1] 

机构地区：[1]第四军医大学西京医院病理科,西安710032 [2]第四军医大学唐都医院全军骨肿瘤研究所,西安710038 [3]第四军医大学基础医学部分子生物学教研室,西安710032

出　　处：《第三军医大学学报》2003年第14期1272-1274,共3页Journal of Third Military Medical University

基　　金：全军"九五"重点补充课题资助项目 ( 98M0 98)

摘　　要：目的　观察分泌型抗肝癌单链双功能抗体基因 (sFv TNF α)的重组逆转录病毒转导的PBMCs对体外培养人肝癌细胞 (HHCC)的杀伤作用。方法　用感染性重组病毒产生细胞C2 2 (PA3 17 PST)产生的病毒上清转导人PBMCs ,采用PCR、RT PCR方法对转导的PBMCs进行DNA和mRNA水平的分析。转导的PBMCs(PBMCs PST)与HHCC共培养 ,MTT法检测PBMCs PST表达产物对肝癌细胞的体外杀伤活性。结果　PCR、RT PCR结果显示PBMCs PST中扩增出外源目的基因对应的电泳条带。MTT法检测结果 ,分泌型抗肝癌单链双功能抗体对体外培养肝癌细胞HHCC的杀伤率为 ( 3 1.4± 4.1) %。结论　分泌型抗肝癌单链双功能抗体基因可以在PBMCs中整合并稳定表达 ,其分泌的表达产物对HHCC具有一定的体外杀伤作用。Objective To investigate the selective cytotoxicity of the bifunctional antibody TNF α fusion proteins in cell line HHCC. Methods The supernatant of the virus produced by C 22 cells generated by transduction using the recombinant retroviral virus was transduced into human peripheral blood mononuclear cells(PBMCs). PCR and RT PCR were used to detect integration and transcription of the sFv TNF α gene in the transduced PBMCs(PBMCs/PST). MTT method was used to detect antitumour activity of the antibody TNF α fusion proteins. Results The results indicated that there was integrated sFv TNF α gene in the genome of PBMCs/PST, and that PBMCs/PST were able to express the fusion antibody TNF α proteins. Cell killing was significant in HHCC cells co cultivated with PBMCs/PST, whereas the PBMCs/pLXSN control cells had no significant cytotoxicity effects on HHCC cells. Conclusion The expression of the bifunctional antibody TNF α fusion proteins in PBMCs/PST has cytotoxicity to HHCC cells in vitro .

关 键 词：肝细胞癌 单链抗体 外周血单个核细胞 体外培养 

分 类 号：R394.2[医药卫生—医学遗传学] R459.9[医药卫生—基础医学]