稳定高效表达TNF-α基因及其突变体的H22肿瘤细胞株的建立  

Establishment of tumor cell line H22 expressing TNF α gene and its mutants with stabitity and highefficacy

作　　者：李清芬[1] 李卓娅[1] 龚非力[1] 姜晓丹[1] 徐勇[1] 冯玮[1] 熊平[1] 

机构地区：[1]华中科技大学同济医学院免疫学教研室,湖北武汉430030

出　　处：《细胞与分子免疫学杂志》2001年第3期210-213,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金重点项目资助;No.39270314

摘　　要：目的建立转染3种TNF-α基因(分泌型TNF-α突变体、跨膜型TNF-α突变体和野生型TNF-α的H22细胞株,以比较不同类型TNF-α的杀瘤效应。方法采用逆转录病毒载体,将3种TNF-α基因导入小鼠H22肿瘤细胞中,用Southern印迹、RT-PCR、流式细胞术和生物学活性检测等方法,从基因的整合、RNA转录、蛋白的表达及其生物学活性4个方面,对转染细胞进行检测鉴定。结果3种TNF-α基因均在H22细胞中得到有效表达,并具有生物学活性。结论获得3种稳定高效表达TNF-α的H22肿瘤细胞株,为进一步研究跨膜型和分泌型TNF-α体内的抑瘤效应建立了良好的实验模型。Aim To establish tumor cell line H22 transfected by TNF α gene and its mutants and compare the tumorocidal effects of different types of TNF α. Methods Mouse hepatocarcinoma cell line H22 were transfected separately with three types of retrovirus vector containing wild TNF α gene and mutants of transmembrane TNF α or secretory TNF α. The transfected cells were further identified for TNF gene integration, RNA transcription, TNF α expression and its activity by Southern blot, RT PCR, FACS and biological activity assay. Results The H22 cells transfected with the three types of TNF α genes could express high levels of TNF α and its mutants, all possessing biological activities. Conclusion The establishment of tumor cell line H22 expressing TNF α gene and its two mutants provide a good model for further studying tumorcidal actions of transmembrane TNF α and secretory TNF α.

关 键 词：跨膜型TNF-Α 分泌型TNF-Α 逆转录病毒载体 肿瘤 H22细胞 细胞毒效应 

分 类 号：R730.51[医药卫生—肿瘤]