嵌合锚定T淋巴细胞对肝癌细胞增殖的阻滞效应  

Repression Effects of Anchor Chemric T Lymphocytes on Proliferation of Tumor Associated Glycoprotein 72 Positive Hepatocarcinoma Cells

作　　者：徐宏勇[1] 徐立[2] 李开宗[3] 

机构地区：[1]解放军第451医院肝胆普外科,陕西西安710054 [2]第四军医大学西京医院消化病研究所,陕西西安710032 [3]第四军医大学西京医院肝胆外科,陕西西安710032

出　　处：《中国普外基础与临床杂志》2011年第4期370-373,共4页Chinese Journal of Bases and Clinics In General Surgery

基　　金：国家自然科学基金项目(项目编号:39900068);陕西省自然科学基金项目(项目编号:2007C259)~~

摘　　要：目的探讨肿瘤相关糖蛋白72(TAG72)靶向性嵌合锚定T淋巴细胞的制备方法,并检测它对TAG72阳性肝癌细胞增殖的阻滞效应。方法分离外周血单核细胞(PBMC),然后用免疫磁珠法分离得到CD8+T淋巴细胞。将重组真核表达载体anti-TAG72-scFv-CD28-pcDNA3.0采用脂质体介导的细胞转染和细胞培养,以制备TAG72靶向性的嵌合锚定T淋巴细胞;将嵌合锚定T淋巴细胞与TAG72阳性肝癌细胞SMMC7721共培养,通过流式细胞仪检测肝癌细胞的周期变化,分析嵌合锚定T淋巴细胞对肝癌细胞增殖的抑制效应。结果 TAG72靶向性嵌合锚定T淋巴细胞可识别肝癌细胞SMMC7721;用流式细胞仪检测发现,嵌合锚定T淋巴细胞可引起肝癌细胞SMMC7721的增殖阻滞。结论 TAG72靶向性嵌合锚定T淋巴细胞可特异性识别TAG72阳性肝癌细胞SMMC7721并引起其增殖阻滞。Objective To investigate the method for generating anchor chemric T lymphocytes that can target tumor associated glycoprotein-72 （TAG72） antigen and analyze their repressive effects on proliferation of TAG72 positive hepatoearcinoma cells. Methods Firstly, peripheral blood mononuelear cells （PBMCs） from healthy vohm reefs were isolated. And then, CD8^＋ T cells were isolated from PBMCs via magnetic activated cell sorting （MACS）. These lymphocytes were transfected with recombinant vector, anti-TAG72-scFv-CD28-pcDNA3, through Lipofectamine2000 to gernerate anchor chimeric TAG72-specific CD8＋ T cells. SMMC7721 （TAG72 positive） hepa- tocarcinoma cells were co-cultured with chimeric T lymphocytes and their cell cycles were analyzed by flow cytome- try （FCM）. Results Anchor chmeric T lymphcytes targetting TAG72 recognized TAG72 positive SMM7721 cells and repressive effects on their proliferation were observed by flow cytometry. Conclusion Anchor chmeric T lymphcytes targetting TAG72 on tumor surface can specifically recognize TAG72 positive hepatocarcinoma cells and may exert repressive effect on their proliferation.

关 键 词：肝癌 肿瘤相关糖蛋白72抗原 单链抗体片段 T淋巴细胞 

分 类 号：R735.7[医药卫生—肿瘤]