一氧化氮诱导HL60细胞凋亡及其机制探讨  

Nitric oxide induced apoptosis in HL-60 cell line and its molecular mechanisms

作　　者：周永列[1] 邱莲女[1] 王文松[1] 皇甫梅生[1] 应奇峰[1] 吴建国[1] 刘建栋[1] 

机构地区：[1]浙江省人民医院,杭州310014

出　　处：《中国免疫学杂志》2004年第2期87-90,共4页Chinese Journal of Immunology

基　　金：浙江省医学科研基金资助 (2 0 0 2A0 0 10 )

摘　　要：目的 :观察一氧化氮对人类白血病细胞是否具有致凋亡作用 ,并研究Bcl 2基因和P5 3基因在此过程中的作用。方法 :将不同浓度的外源性一氧化氮供体亚硝基铁氰化钠与HL 6 0细胞作用 ,观察其作用的时间效应和剂量效应 ;用MTT法观察NO对细胞的抑制作用 ,用透射电镜观察细胞结构改变 ,用DNA凝胶电泳、细胞DNA含量、DNA末端标记、Annexin V PI法等分析细胞凋亡 ,并用流式细胞法进一步观察在NO作用过程中凋亡调控因子Bcl 2和P5 3蛋白及线粒体膜蛋白表达变化。结果 :NO能抑制HL 6 0细胞生长 ,并在一定的剂量范围内显示作用时间和剂量的量效关系 ;典型的细胞形态改变、DNA片段化、亚G1峰检出、DNA末端标记、Annexin V PT- 表达增加等证实NO能诱导白血病细胞凋亡 ;在此过程中 ,P5 3蛋白表达上调 ,而Bcl 2蛋白表达下调 ,线粒体膜蛋白表达增加。结论 :NO对HL 6 0细胞有强的致凋亡作用 ,Bcl 2和P5 3参与NO诱导HL 6 0细胞凋亡的调控。Objective:To observe whether nitric oxide(NO) can induce apoptosis in HL-60 cell line and further study the role of Bcl-2 gene and P53 gene in this process.Methods:Different concentrations of NO were used to treat HL-60 cell. The growth inhibition was analysed by MTT assay. Cell morphology was observed under transmission electromieromicroscope. Cell apoptosis was analysed by DNA agarose gel electrophoresis, DNA content, TUNEL, and Annexin-V/PI labeling method. Bcl-2 and P53 gene proteins and mitochondrie membrane protein by Flow cytometry. Results:NO can inhibit HL-60 cell growth. Within a certain range of treating time and does, cell were induced apoptosis with a time and does related manner. Cell apoposis was comfirmed by type cell morphology and DNA fragment and sub-G1 phase and TUNEL and Annexin-V/PI labeling method. The expressinos of P53 protein and mitochondrial membrane protein were increased and Bcl-2 protein became decreased after NO treatment. Conclusion:NO could efficiency induce apoptosis of HL-60 cell line. Bcl-2 and P53 gene participate in the NO induced apotosis of HL-60 cells.

关 键 词：一氧化氮 细胞株 HL-60 细胞凋亡 BEL-2基因 T53基因 

分 类 号：R55[医药卫生—血液循环系统疾病]