低浓度哇巴因对人白血病细胞株Jurkat的生长调控作用及其分子机制初步研究  

Effects and Mechanism of Ouabain at Low Concentrations on the Jurkat Cells Growth

作　　者：杨磊[1] 郭雪梅[1] 马玉娟[1] 李娟[1] 欧阳建[1] 

机构地区：[1]南京大学附属鼓楼医院血液科,江苏南京210008

出　　处：《现代生物医学进展》2012年第13期2436-2439,共4页Progress in Modern Biomedicine

基　　金：国家自然基金资助项目(30971511);人事部留学回国人员启动基金(2009);教育部留学回国人员启动基金(41批)

摘　　要：目的:观察低浓度哇巴因对人白血病细胞株Jurkat生长的影响并初步探讨哇巴因特异性调节Jurkat细胞生长的机制。方法:分别用不同低浓度哇巴因作用于人白血病细胞株Jurkat后,采用四甲基偶氮唑盐MTT法检测细胞增殖情况、流式细胞学FCM技术检测细胞凋亡情况以及细胞内线粒体膜电位变化情况,Western blot法观察哇巴因对Jutkat细胞膜表面钠钾ATP酶的表达调节作用。结果:MTT及FCM检测结果表明随着哇巴因浓度的增高,哇巴因对Jurkat细胞的增殖抑制及促凋亡作用越明显。WesternBlot结果显示30nM及50nM哇巴因作用于Jurkat细胞株48h后引起细胞钠钾ATP酶表达下调,[3H]-哇巴因结合实验结果显示在Jurkat细胞株随着哇巴因作用浓度升高,细胞膜钠泵对哇巴因的亲和力逐渐下降。结论:低浓度哇巴因即可抑制白血病细胞株Jurkat增殖并诱导其凋亡。这种特异性细胞生长调控作用与哇巴因引起的细胞膜钠钾ATP酶表达变化相关,最终引起细胞内线粒体膜电位发生变化,释放相关凋亡蛋白,诱导细胞凋亡。Objective： To investigate the effect and the mechanism of ouabain at low concentrations on the growth in the Jurkat leukemia cell lines.Methods： Cell proliferation was observed by using MTT method.The cell apoptosis and the mitochondrial membrane potential changes in cells were assayed by Streaming cytology technology.The expressions of Na/K-ATPase Subunit were evaluated by western-blot before and after the cells were treated with different low concentration ouabain.Results： MTT tests showed that with the increasing of the concentration of ouabain,the proliferation of the Jurkat cell suppressed and the apoptosis role increasing.The result of western-blot showed that low concentrations of ouabain（30nM 50nM） caused Na/K-ATPase expression fall on the Jurkat cell for 48h.The -ouabain combining tests showed that with the increasing of the concentration,of ouabain the active Na/K-ATPase in the plasma membrane gradually decreased.Conclusions： The low concentration ouabain can not only inhibit the proliferation of the Jurkat cells,but also induce its apoptosis.This effect and the Na/K-ATPase expression changes related.That changes caused the mitochondrial membrane potential to change finally thus to induce the apoptosis by releasing the apoptosis proteins.

关 键 词：哇巴因 钠钾ATP酶 JURKAT细胞 增殖 凋亡 

分 类 号：Q75[生物学—分子生物学] Q78