多胺缀合物WJH-6诱导白血病细胞凋亡机制研究

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Western blotting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。

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

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.