亚硒酸钠对HL-60细胞增殖、凋亡影响及作用机制探讨  

The effects and mechnanisms of sodium selenite on inducing HL60 cells apoptosisand growth inhibition

作　　者：贾永清[1] 滕熔[2] 胡慧仙[1] 

机构地区：[1]金华市中心医院血液科,浙江321000 [2]南通大学附属医院血液科

出　　处：《交通医学》2011年第2期121-125,共5页Medical Journal of Communications

摘　　要：目的:以人急性髓细胞性白血病细胞系HL-60为模型,探讨亚硒酸钠对HL-60细胞增殖及凋亡的影响。方法:以不同浓度的亚硒酸钠作用于HL-60细胞后,分别采用台盼蓝拒染法计数活细胞,MTT法检测细胞增殖抑制;通过光镜及双染流式细胞检测分析细胞凋亡;检测线粒体膜电位变化。分光光度法检测caspase-3的活性变化,RT-PCR检测Bcl-2的mRNA的表达。结果:(1)用台盼蓝染色和MTT方法证实亚硒酸钠(>5μmol/L)能抑制HL-60细胞的生长及存活。(2)HL-60细胞经亚硒酸钠处理后,形态学上出现凋亡细胞特征性改变,双染流式细胞检测证实,亚硒酸钠能有效诱导HL-60细胞凋亡。(3)亚硒酸钠处理HL-60细胞后,流式细胞仪检测线粒体膜电位下降,分光光度法显示caspase-3活性升高,凋亡基因Bcl-2的mRNA的表达明显降低。结论:亚硒酸钠能有效抑制HL-60细胞的增殖及存活,且能够诱导其凋亡,抑制率及凋亡率与药物剂量呈相关。线粒体介导的凋亡途径是亚硒酸钠诱导HL-60细胞凋亡的可能机制。Objective： To use human myeloid leukemia cell line HL-60 as an experimental model,to research effects of sodium selenite on inducing HL-60 cells apoptosis and growth inhibition.Methods：After action on HL-60 cells in different densities of sodium selenite,to calculate the numbers of living cells by using typan blue exclusion test,inhibition rate was tested by MTT method.Apoptosis induced by sodium selenite in HL-60 cells was investigated by applying Light microscopy and flow cytometry Annexin-V/PI double staining analysis.Flow cytometry analysis was used to exam mitochondrion membrane potential;Colorimetric method was used to measure the activities of caspase-3 of HL-60;RT-PCR was employed to analyze the mRNA expression levels of Bcl-2.Results：（1）Sodium selenite was found to be able to inhibit the proliferation and survival of HL60 cells by typan blue staining and MTT assay.（2）After dealing with HL60 cells in sodium selenite,classic character of morphologic change of apoptotic cells was observed with an inverted microscope.It was showed that sodium selenite was able to induce HL60 cell apoptosis by staining with Annexin V.（3）With different densities of sodium selenite in HL60 cells,mitochondrion membrane potential was cut down by FCM,activities of caspase3 were heightened by using colorimeteic method,and the mRNA expression levels of Bcl-2 became lower.Conclusion：Sodium selenite may effectually inhibit multiplication and survival of HL60,to induce its apoptosis.Mitochondrion-mediated apoptotic cell death way was possible apoptotic mechanism of HL60 cells.

关 键 词：急性髓细胞性白血病 亚硒酸钠 HL-60细胞 凋亡 线粒体膜电位 caspase-3 Bcl-2 流式细胞仪检测 逆转录聚合酶链反应 

分 类 号：R733.7[医药卫生—肿瘤]