bcl-2基因转染阻断Fas介导的Jurkat细胞凋亡  

Fas mediated apoptosis inhibited by human bcl-2 gene in lymphoma cell line Jurkat

作　　者：曹惠芳[1] 钱其军[1] 吴孟超[1] 黄洪莲 王华菁[1] 贾随旺 郭亚军[1] 

机构地区：[1]上海第二军医大学东方肝胆外科医院肿瘤免疫和基因治疗中心

出　　处：《中华血液学杂志》1998年第5期244-246,共3页Chinese Journal of Hematology

基　　金：国家自然科学基金;国家自然科学基金;国家及上海市博士后基金

摘　　要：目的：通过ｂｃｌ２阻断Ｆａｓ介导的细胞凋亡了解ｂｃｌ２基因在淋巴瘤癌变过程中逃避机体免疫监控的作用机制。方法：应用基因重组技术重组ｐＬＸＳＮｂｃｌ２，采用电穿孔法将其与空载体分别转染包装细胞系ＰＡ３１７，将阳性克隆病毒上清感染人Ｔ淋巴瘤细胞株Ｊｕｒｋａｔ，用Ｇ４１８筛选，其阳性克隆进行免疫组化检测，应用抗Ｆａｓ抗体诱导细胞凋亡来模拟细胞毒Ｔ细胞的杀伤机制，观察ｂｃｌ２在淋巴瘤逃避免疫机制中的作用。结果：转染人ｂｃｌ２基因的Ｊｕｒｋａｔ（Ｊｕｒｋａｔｂｃｌ２）细胞较对照Ｊｕｒｋａｔ及转染空载体Ｊｕｒｋａｔ（Ｊｕｒｋａｔｎｅｏ）细胞，ｂｃｌ２表达量明显增加，而Ｆａｓ基因表达量无明显变化。其Ｊｕｒｋａｔｂｃｌ２能明显耐受抗Ｆａｓ抗体诱导的细胞凋亡。结论：人ｂｃｌ２基因在淋巴瘤中过量表达，能部分阻断抗Ｆａｓ抗体诱导的细胞凋亡，ｂｃｌ２基因过量表达可能是淋巴瘤癌变过程中逃脱机体免疫监控的机制之一。Objective: To understand the mechanism of bcl-2 gene in escaping the immune surveillance in the development of lymphoma. Methods: A recombinant retroviral vector pLXSNbcl-2 was constructed by cloning bcl-2 cDNA into the replication defective retroviral vector pLXSN, and transferred to packaging cell line PA317 by electroporation. The G418 resistant colonies were selected, and the supernatants of the colony cultures were used to infect the human lymphoma cell line Jurkat. Cells in G418 resistant Jurkat colonies were characterized by immunohistochemistry. Anti-Fas monoclonal antibody (McAb) was applied to Jurkat cells for inducing apoptosis which mimicked the cytotoxic activity of T lymphocyte. Results: Expression of bcl-2 in pLXSNbcl-2 transfected Jurkat cell (Jurkatbcl-2) increased, while there was no change of Fas gene expression. Apoptosis was blocked in Jurkatbcl-2 by anti-Fas McAb treatment. Conclusion:Overexpression of bcl2 in lymphoma cell line could inhibit cell apoptosis induced by anti Fas McAb, suggesting that overexpression of bcl-2 is one of the mechanisms in escaping immune surveillance in the development of lymphoma.

关 键 词：淋巴瘤 细胞凋亡 BCL-2 基因转染 JURKAT细胞 

分 类 号：R733[医药卫生—肿瘤]