跨膜型肿瘤坏死因子信号肽氨基酸部分缺失对细胞毒作用影响  

Deletions of different region of human transmembrane tumor necrosis factor (TM-TNF-α) signal peptide and cytotoxic activity

作　　者：姜小丹[1] 李清芬[1] 龚非力[1] 徐勇[1] 冯玮[1] 熊平[1] 李卓娅[1] 

机构地区：[1]华中科技大学同济医学院免疫教研室,武汉430030

出　　处：《中华微生物学和免疫学杂志》2004年第7期530-532,共3页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目 (批准号 :3 9870 713 )

摘　　要：目的　探讨跨膜型肿瘤坏死因子 (TM TNF)信号肽在其杀瘤中的作用。方法　用PCR构建突变体及体外转录、翻译蛋白质 ,分别获得TM TNF信号肽胞浆段、胞外与分泌型肿瘤坏死因子(S TNF)连接段以及部分跨膜段缺失即信号肽第 73～ 4 7、 73～ 34、 34～ 1、 2 1～ 1位氨基酸缺失的TM TNF体外翻译蛋白。用MTT法检测其对HL 6 0细胞的杀伤率。结果　 34～ 1、 2 1～ 1位氨基酸缺失可提高TM TNF对HL 6 0细胞的杀伤 , 73～ 4 7位氨基酸缺失对其杀瘤力无明显作用 ,而 73～ 34位氨基酸缺失可使其杀瘤率明显下降。结论　去除信号肽胞外段可提高TM TNF α对HL 6 0细胞的杀伤率 ,而TM TNF α信号肽 4 7～ 34位氨基酸在其杀瘤过程中有重要作用。Objective To study the effects of a series of TM-TNF-α signal peptide mutations with deletions occurring the region covering cytoplasmic transmembrane and linking domains (-1 to -21,-34 and -73 to -49,-34) on the cytotoxic activity against HL-60. Methods The proteins of four deletions region of TM-TNF signal peptide were produced by in vitro transcription and translation system in a microsomal. The mutant DNA fragment was obtained by recombinant PCR. Results Four recombinant proteins were generated and assayed for HL-60 cytotoxic activity with MTT. The killing rates against HL-60 of △-73 to -47,△-21 to -1,△-32 to -1,△-73 to -34 TM-TNF and W-TNF are 40.9%,55.58%,54.47%,18.25%,41.5% respectively. Conclusion It is the deletion of linking domains of TM-TNF -47 to -34 that decreases cytotoxic activity of TM-TNF against HL-60.

关 键 词：跨膜型 肿瘤坏死因子 信号肽 氨基酸 细胞毒作用 TM-TNF PCR 

分 类 号：R392[医药卫生—免疫学]