论文部分内容阅读
目的探讨嘌呤能Ｐ２ｚ受体在介导慢性淋巴细胞白血病（ＣＬＬ）细胞凋亡中的作用。方法将表达Ｐ２ｚ受体［Ｐ２ｚ（＋）］与不含Ｐ２ｚ受体［Ｐ２ｚ（－）］两组ＣＬＬ细胞分别同１．０ｍｍｏｌ／Ｌ三磷酸腺苷（ＡＴＰ）体外培养８小时，以电镜、ＤＮＡ凝胶电泳和定量ＤＮＡ３′－末端的ＴｄＴ法检测细胞凋亡；并对ＡＴＰ、ＢｚＡＴＰ、２ＭｅＳＡＴＰ、ＡＴＰ－γＳ与其他核苷的诱导及ＯｘＡＴＰ、ＫＮ－６２的抑制效应做定量研究。结果（１）Ｐ２ｚ（＋）细胞能在ＡＴＰ诱导下呈现典型的细胞凋亡形态和ＤＮＡ梯状条带；（２）不同的Ｐ２ｚ受体激活剂可通过Ｐ２ｚ受体诱导细胞凋亡并产生不同量的ＤＮＡ降解片段，诱导能力的强弱顺序是ＢｚＡＴＰ＞ＡＴＰ＞２ＭｅＳＡＴＰ，而ＡＴＰ－γＳ或其他核苷几乎无诱导能力；（３）Ｐ２ｚ受体的抑制剂Ｏｘ－ＡＴＰ和钙调节蛋白激酶抑制剂ＫＮ－６２可完全阻止诱导剂诱导的细胞凋亡的诱导。结论Ｐ２ｚ受体在介导由ＡＴＰ诱导ＣＬＬ细胞凋亡的过程中起着十分重要的作用。
Objective To investigate the role of purinergic P2z receptors in mediating apoptosis of chronic lymphocytic leukemia (CLL) cells. Methods CLL cells expressing P2z receptor [P2z(+)] and P2z receptor-free [P2z(-)] were cultured with 1.0 mmol/L adenosine triphosphate (ATP) in vitro for 8 hours, with electron microscope and DNA gel. Electrophoresis and quantitative DNA 3′-end TdT assay was used to detect apoptosis; quantitative studies were performed on the induction of ATP, BzATP, 2MeSATP, ATP-γS and other nucleosides, and the inhibitory effects of OxATP and KN-62. Results (1) P2z(+) cells can exhibit typical apoptotic morphology and DNA ladders induced by ATP; (2) different P2z receptor activators can induce apoptosis and produce differences through P2z receptors. The order of the inducing ability of the amount of DNA degrading fragment is BzATP>ATP>2MeSATP, while ATP-γS or other nucleosides have almost no inducing ability; (3) P2z receptor inhibitor Ox-ATP and calcium-regulated protein kinase inhibition Agent KN-62 can completely prevent inducer-induced apoptosis induction. Conclusion The P2z receptor plays an important role in mediating the apoptosis of CLL cells induced by ATP.