多胺缀合物WJH-6诱导白血病细胞凋亡机制研究

Apoptotic mechanism of WJH-6,a novel polyamine conjugate,on K562 and HL-60 cells

探讨新型多胺缀合物WJH-6对多胺转运体的识别及其诱导K562、HL-60白血病细胞凋亡的机制。采用MTT法检测细胞毒性;应用流式细胞仪检测细胞周期及凋亡率的变化;应用高内涵活细胞成像系统检测WJH-6对多胺转运体的识别,细胞内含量的变化及对线粒体膜电位、Bid、Caspase-3、-8、-9的影响;采用Westernblotting方法检测线粒体及胞浆中细胞色素c含量的变化。实验结果显示,WJH-6具有良好的多胺转运体识别能力,并可诱导白血病K562、HL-60细胞线粒体膜电位降低、细胞色素c释放以及Bid、Caspase-3、-8、-9等活化。本研究结果提示,WJH-6诱导的白血病K562、HL-60细胞凋亡与线粒体损伤有关。

In the present study,the apoptotic mechanism and polyamine transporter recognition of WJH-6,a novel polyamine conjugate,were investigated in K562 and HL-60 cells. The cytotoxicity of WJH-6 was assessed by MTT assay; cell cycle distribution and apoptosis were measured by flow cytometry; the protein expression of Caspase-3,Caspase-8,Caspase-9,Bid and mitochondrial membrane potential （MMP） were evaluated by high content screening （HCS） analysis; the protein expression of cytochrome c was measured by Western blotting. The results showed that WJH-6 could be recognized and transported by polyamine transporter （PAT）. Furthermore,WJH-6 was able to inhibit K562 and HL-60 cells proliferation and induce apoptosis. This apoptotic effect was relative to MMP loss,cytochrome c release from mitochondria to cytoplasm and the activation of Caspase-8,Caspase-9,Caspase-3 and Bid. These results suggested that WJH-6-induced K562 and HL-60 cells apoptosis was related with mitochondrial damage.