Bcl-2,Bax和线粒体膜蛋白在一氧化氮供体诱导HL-60细胞凋亡中的改变

Change of Bcl-2,Bax Proteins and Mitochondrial Membrance Protein on Nitric Oxide Induced Apoptosis in HL-60 Cells

本研究探讨外源性一氧化氮 (NO)供体硝普钠 (SNP)对HL 6 0细胞诱导凋亡的可能机制。将HL 6 0细胞与SNP在体外培养 ,用DNA片段原位末端标记法 (TUNEL)测定原位细胞凋亡率 ;用流式细胞仪测定细胞DNA倍体和周期分析及Bcl 2、Bax、线粒体膜蛋白表达率的变化。结果表明 :SNP可诱导HL 6 0细胞凋亡 ,两者之间有明显的量效和时效关系。 1.0mmol/LSNP作用 4 8小时后 ,HL 6 0细胞的凋亡率分别为亚二倍体峰 (4 2 .2± 3.5 ) % ,TUNEL测定凋亡细胞率为 (5 2 .5± 7.6 ) % ,显著高于空白对照组和同浓度的高铁氰化钾 (PFC)组 ;Bax基因蛋白和线粒体膜蛋白 (APO2 .7)表达增加 ,bcl 2基因蛋白表达降低。Bax、Bcl 2和APO2 .7的表达率与SNP两者之间也有明显的量效和时效关系。结论 :外源性一氧化氮供体诱导HL 6 0细胞凋亡过程中 ,线粒体膜蛋白表达显著上调并伴随Bax和Bcl 2蛋白的表达改变。

To investigate the possible mechanisms of nitric oxide(NO)-induced apoptosis in leukemia cell line HL-60,HL-60 cells in vitro were incubated with sodium nitroprusside (SNP),the in situ cell apoptosis quantitatively was assayed by TdT-mediated dUTP nick end labeling (TUNEL),the cell cycle DNA and proteins expression of Bcl-2,Bax,mitochondrial membrance protein (APO2.7) were analyzed by flow cytometry. The results showed that SNP induced HL-60 cell apoptosis in a dosage- and time- dependent manner. After exposure to SNP at the concentration of 1.0 mmol/L for 48 hours,the percentage of apoptosis HL-60 was (42.2±3.5)% for subG1 and (52.5±7.6)% for TUNEL respectively,and they are significantly higher than those in control and potassium ferricyanide (PFC) groups as same concentration. During the apoptosis process,it showed a decrease of Bcl-2 protein and an increase of Bax protein and mitochondrial membrance protein in HL-60 cell,proteins of Bcl-2,Bax and mitochondrial membrance were expressed in a dosage- and time-dependent manner too. In conclusion,during the process of SNP induced apoptosis in HL-60 cell,the expression of mitochondrial membrane protein was increased,Bcl-2 and Bax proteins may be important regulators.