摘要
目的 研究可溶性人TRAIL蛋白 (sTRAIL)对Jurkat细胞的生长抑制效应及凋亡诱导作用。方法 通过RT PCR扩增人TRAIL分子胞外区 4 1～ 2 81的密码子 ,利用大肠杆菌DH5α表达该可溶性片段 ,经纯化和复性后 ,诱导Jurkat细胞 ,通过显微镜、台盼蓝排斥试验、MTT法、流式细胞仪和DNA断裂实验检测细胞增殖和细胞凋亡。结果 sTRAIL蛋白纯度达 90 %以上。用 0 .5～ 10 .0 μg/ml的蛋白诱导Jurkat细胞 12h以上细胞的生长和增殖即被显著抑制 ,并且观察到凋亡峰及DNA的片段化等凋亡的特征性变化 ,细胞的生长抑制率与凋亡率也呈剂量依赖和时间依赖关系。结论 利用大肠杆菌制备的sTRAIL41 2 81蛋白对Jurkat细胞具有明显的增殖抑制效应和凋亡诱导作用。
Objective To investigate the effects of human so luble TRAIL(sTRAIL) on growth and apoptosis of Jurkat cells. Method s The cDNA encoding human TRAIL extracellular region(amino acids 41 -281) was amplified by RT-PCR, the soluble segment was expressed in E.coli DH5α, purified and refolded. After treatment of sTRAIL, the cell proliferation and apoptosis were determined by morphological approach; MTT assay, flow cytome try and DNA fragmentation assay. Results Homogeneity of h arvested sTRAIL was more than 90%. Proliferation of Jurkat cells was significant ly inhibited 12 h after the addition of 0.5-10.0μg/ml sTRAIL. Apoptosis peak(A P) and DNA ladder were observed. The effect of sTRAIL on cell proliferation and apoptosis depended on the concentration and the time of incubation. C onclusion The purified and refolded sTRAIL 41-281 expressed by E.coli inhibited the growth of Jurkat cells and induced apoptosis. [
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2004年第9期682-686,共5页
Chinese Journal of Microbiology and Immunology
