死亡受体5诱导Jurkat细胞凋亡的作用论文
2015-04-24 01:01
导读：药学论文毕业论文，死亡受体5诱导Jurkat细胞凋亡的作用论文怎么写，格式要求，写法技巧，科教论文网展示的这篇论文是很好的参考：
【关键词】 细胞凋亡 Role of DR5 in TRAIL induced apoptosis of Jurkat
【关键词】 细胞凋亡
Role of DR5 in TRAIL induced apoptosis of Jurkat cells
【Abstract】 AIM: To study the role of DR5 in TRAIL apoptotic signal transduction in Jurkat cells. METHODS: cDNA containing fulllength extracellular domain of human DR5 was cloned into pGAPZα. Recombinant Pichia pastoris clone generated via homologous recombination secreted high levels of sDR5. BALB/c mice were immunized with sDR5 in CFA to prepare antiDR5 mAb. The binding of antiDR5 mAb was measured for surface expression of TRAIL receptor by flow cytometric analysis. Jurkat cells were tested for their susceptibility to apoptosis induced by TRAIL with TRAIL apoptosis kit so as to study the correlation between DR5 expression level and TRAIL sensitivity. The level of DR5 expression on Jurkat cell line was examined by flow cytometry. The rates of TRAILinduced apoptosis and antiDR5 mAb blocking on Jurkat cells were tested by flow cytometry with TRAIL apoptosis kit. RESULTS: The killing role of TRAIL was blocked in Jurkat cells pretreated with antiDR5 mAb. The average percentage of blocking was 90.49%. CONCLUSION: AntiDR5 mAb can specifically bind to DR5 and DR5 is expressed at high levels on Jurkat cells. DR5 plays a very key role in TRAIL induced apoptosis in Jurkat cells. DR5 expression is important for the induction of apoptosis by TRAIL.
【Keywords】 Jurkat cells; TRAIL; apoptosis; DR5; antibodies, monoclonal
【摘要】 目的： 探讨DR5在TRAIL诱导Jurkat细胞凋亡中的作用. 方法： 用含人全长DR5细胞外全长结构域重组DR5免疫BALB/c小鼠，制备抗DR5 mAb. 将96孔U型细胞培养板加1×105/孔Jurkat细胞和5 mg/L抗DR5 mAb，采用TRAIL试剂盒检测细胞凋亡率. 结果： 抗DR5 mAb与Jurkat细胞表面的DR5作用后，TRAIL对Jurkat细胞的杀伤功能几乎被抗DR5 mAb完全阻断，阻断率高达90.49%. 结论： DR5在TRAL诱导的Jurkat细胞凋亡中起关键作用.
【关键词】 Jurkat细胞；TRAIL；细胞凋亡；DR5；抗体，单克隆
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0引言
肿瘤坏死因子相关凋亡诱导配体（tumor necrosis factor related apoptosis inducing ligand, TRAIL）是近几年新发现的TNF 超家族成员，TRAIL与其受体结合可以诱导大多数肿瘤细胞凋亡而对大多数正常细胞无影响. 已经发现，TRAIL有5种受体，即2种死亡受体（DR4, DR5）、2种诱骗受体（DcR1, DcR2）及可溶性受体OPG(osteoprotegerin). 其中只有DR4［1］和或DR5有胞内死亡结构域，诱导细胞凋亡. DR4和DR5在诱导细胞凋亡方面具有相同的作用，但哪一种受体在TRAIL诱导肿瘤细胞凋亡方面更为重要仍不清楚. 发展和应用TRAIL 不同受体的特异性抗体是分析这两种不同受体所参与的死亡途径的关键. 我们用特异性抗DR5 mAb阻断DR5的功能，并应用流式细胞仪检测TRAIL诱导Jurkat细胞凋亡情况，以探讨DR5在TRAIL诱导肿瘤细胞凋亡中的作用.
1材料和方法
1.1材料
含有人DR5细胞外结构域全长cDNA的毕赤酵母表达载体菌株由美国宾夕法尼亚大学医学院病理医学实验室陈有海教授馈赠. Jurkat细胞为本室保存细胞株传代培养于含2.0 mmol/L L谷氨酰胺、1.5 g/L碳酸氢钠、10.0 mmol/L HEPES, 1.0 mmol/L丙酮酸钠、100.0 mL/L FBS, 1×105 U/L 青霉素和100 mg/L链霉素的RPMI1640培养基. DMEM/HAT和DMEM/HT选择培养基（Sigma公司）. TRAIL凋亡检测试剂盒（Upstate biotech）. FACSCalibur型流式细胞仪（美国BD公司）.
1.2方法
1.2.1抗DR5 mAb制备及特异性鉴定
