转导单链免疫毒素基因的PBMCs对人肝癌细胞SMMC-7721的体外杀伤活性  

In vitro cytotoxicity of PBMCs via genetic modification of single-chain immunotoxin

作　　者：程虹[1] 刘彦仿[1] 张惠中[2] 沈万安[2] 张菊[3] 张静[1] 

机构地区：[1]中国人民解放军第四军医大学基础部病理学教研室,陕西省西安市710033 [2]中国人民解放军第四军医大学唐都医院全军骨肿瘤研究所,陕西省西安市710038 [3]中国人民解放军第四军医大学基础部生物化学教研室,陕西省西安市710033

出　　处：《世界华人消化杂志》2003年第3期281-284,共4页World Chinese Journal of Digestology

基　　金：军队"九.五"重点补充课题;No.98M098~~

摘　　要：目的:用携带分泌型抗肝癌单链免疫毒素基因(sFv-TNF-α)的重组逆转录病毒感染人外周血单个核细胞(peripheralbloodmononuclearcells,PBMCs),使其表达并分泌针对人肝癌细胞的sFv-TNF-α融合蛋白,观察转导的PBMCs对体外培养人肝癌细胞SMMC-7721的杀伤作用.方法:用感染性重组病毒产生细胞C22(PA317/PST)产生的病毒上清转导人PBMCs,采用PCR和RT-PCR方法对转导的PBMCs(PBMCs/PST)进行DNA和mRNA水平的分析.PBMCs/PST与SMMC-7721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及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,分泌型抗肝癌单链免疫毒素对体外培养肝癌细胞SMMC-7721的杀伤率为(38.2±6.7)%.结论:分泌型抗肝癌单链免疫毒素基因可以在PBMCs中整合并稳定表达,其分泌的表达产物对SMMC-7721具有一定的体外杀伤作用.AIM:To investigate the selective cytotoxicity of single-chain immunotoxin (sFv-TNF- α fusion proteins) in cell line SMMC-7721. METHODS:HCC-specific killer cells were generated by transducing the recombinant retroviral virus in supernatant of the virus producing cells 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 transduced PBMCs (PBMCs/PST). MTT method was used to detect antitumour activity of the sFv-TNF-α fusion proteins. RESULTS:There was integrated sFv-TNF-α gene in the genome of PBMCs/PST, and PBMCs/PST were able to express the fusion sFv-TNF-α proteins. Cell killing was significant in HCC cells co-cultivated with PBMCs/PST, whereas the PBMCs/ pLXSN control cells had no significant cytotoxic effects on HCC cells. CONCLUSION:Expression of sFv-TNF-α fusion proteins in PBMCs/PST has cytotoxicity to HCC cells in vitro.

关 键 词：转导单链免疫毒素基因 PBMCS 肝癌 SMMC-7721 体外杀伤活性 

分 类 号：R735.7[医药卫生—肿瘤] R730.51[医药卫生—临床医学]