一氧化氮诱导HL-60细胞凋亡及其机制探讨

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P53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-60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P53蛋白及线粒体膜蛋白表达变化。结果：NO能抑制HL-60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，P53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-60细胞有强的致凋亡作用，Bcl-2和P53参与NO诱导HL-60细胞凋亡的调控。

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

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.