摘要
本研究旨在探讨线粒体途径在慢性髓系白血病(CML)信号转导中的作用。用脂质体转染法将bcr3/abl2反义寡核苷酸(ASO)导入CML细胞系K562细胞中;用MTT法检测bcr3/abl2ASO对K562细胞活力的影响;流式细胞术(FCM)分析测定细胞凋亡率;荧光染料罗丹明123染色分析细胞线粒体跨膜电位(ΔΨm)的变化;Westernblot检测线粒体凋亡信号转导通路相关蛋白细胞色素C(CytC)的表达。结果表明,2μmol/Lbcr3/abl2ASO与K562细胞作用24小时后,可明显抑制K562细胞活力,其抑制率为65.7%;可诱导细胞凋亡,凋亡率为16.9%;可下调K562细胞线粒体ΔΨm,有38.33%的细胞出现线粒体ΔΨm下降;可增强CytC的表达,激光光度扫描仪测得其表达光密度值由2.33±0.3升高到4.78±0.1。结论:线粒体途径通过介导凋亡信号而在CML信号转导中发挥重要作用。
This study was aimed to investigate the role of mitochondia pathway in signal transduction of chronic myeloid leukemia（CML） . After bcr3/abl2 antisense oligodeoxynucleotide （ASO） was introduced into CML cell line K562 cells by liposomal transfection, the cell viability was detected by MTT assay, the cell apoptosis was determined by flow- cytometry （FCM） , the mitochondrial membrane potential （△ψ） was labeled by Rhodamine123 and examined by FCM, and the expression of mitochondrial apoptosis signal transduction pathway related proteins cytochrome C was ana- lyzed by Western blot. The results showed that after K562 cells were exposed to 2 μmol/L of bcr3/abl2 ASO for 24 hours, bcr3/abl2 ASO significantly inhibited cell viability with inhibitory rate of 65.7%, induced the apoptosis of K562 cell line with apoptotic rate of 16.9%, and decreased mitochondrial △ψ of K562 cells with the reducing rate of 38.33%, enhanced the expression of cytochrome C with increase of optical density value from 2.33 ＋ 0.3 to 4.78＋ 0.1 by laser photometric scanning. It is concluded that mitochondia pathway plays an important role in signal transduction of chronic myeloid leukemia by directing apoptotic signal transduction.
出处
《中国实验血液学杂志》
CAS
CSCD
2009年第2期277-280,共4页
Journal of Experimental Hematology
基金
国家自然科学基金课题资助,编号39670330、39770336
关键词
慢性髓系白血病
线粒体途径
反义寡核苷酸
信号转导
线粒体跨膜电位
chronic myeloid leukemia
mitochondia pathway
antisense oligodeoxynucleotide
signal transduction
mitochondrial membrane potential
