GPI-PLD基因表达在K562细胞对补体杀伤的敏感性中的影响  

Effect of the expression of glycosylphosphatidylinositol-specific phospholipase D gene on K562 cell sensitivity to complement killing

作　　者：王依丹[1] 唐建华[1] 杨智英[1] 禹虹[1] 向新颖[1] 

机构地区：[1]中南大学生物科学与技术学院生物化学系,长沙410078

出　　处：《中南大学学报（医学版）》2004年第6期654-657,共4页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金 (30 2 71 4 56)

摘　　要：目的 :观察葡萄糖、胰岛素诱导K5 6 2细胞GPI PLD基因表达对GPI锚定CD5 5 ,CD5 9的影响 ,以及对补体依赖的细胞毒 (complementdependentcytotoxicity ,CDC)杀伤K5 6 2细胞的影响。方法 :采用免疫印迹检测CD5 5和CD5 9表达 ,定量测定GPI PLD酶活性 ,台盼蓝排斥法观察CDC杀伤率。结果 :诱导 2 4h及 4 8h ,胰岛素组、葡萄糖 +胰岛素组酶活性、杀伤率明显增高。 2 4h时对照组、葡萄糖组、胰岛素组膜蛋白检出CD5 5 ,对照组、葡萄糖组膜蛋白及胰岛素组、葡萄糖 +胰岛素组培养基中检出CD5 9。 4 8h时对照组膜蛋白 ,葡萄糖组、胰岛素组、葡萄糖 +胰岛素组培养基检出CD5 5 ,CD5 9。结论 :胰岛素诱导使K5 6 2细胞的GPI PLD酶活性明显升高 ,导致GPI锚定CD5 5和CD5 9释放进入培养基 ,K5 6Objective To detect the release of glycosylphosphatidylinositol (GPI) anchored CD55 and CD59, and the effect of complement dependent cytotoxicity on K562 cell with expression of GPI-PLD by glucose or insulin. Methods CD55 and CD59 were detected by Western blotting; GPI-PLD activities were analyzed quantitatively; and complement dependent cytotoxicity (CDC) effects were observed by staining of trypan blue. Results After being induced by insulin or glucose for 24 h and 48 h, GPI-PLD activities and rate of CDC killing in the insulin, insulin+glucose groups significantly increased. At 24 h after the inducement, CD55 was found in the membrane proteins in the control, glucose and insulin groups and CD59 in membrane proteins in the control, glucose group, and medium supernatants of insulin, glucose+insulin groups. Both CD55 and CD59 were found in membrane proteins in control group, and medium supernatants of glucose, insulin, glucose+insulin group at 48 h after the inducement.Conclusion Treatment with insulin resulted in the obvious increase of GPI-PLD activity in K562 cell, which led to releasing of GPI anchored CD55 and CD59 into medium,and increased sensitivity of these cells to CDC killing.

关 键 词：胰岛素 CD55 K562细胞 GPI CD59 补体 葡萄糖 膜蛋白 酶活性 基因表达 

分 类 号：R73-3[医药卫生—肿瘤]