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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。
To investigate the recognition of polyamine transporter by novel polyamine conjugate WJH-6 and its mechanism of inducing apoptosis in K562 and HL-60 leukemia cells. The cytotoxicity was detected by MTT assay. The changes of cell cycle and apoptosis rate were detected by flow cytometry. High-content living cell imaging system was used to detect the changes of WJH-6 on polyamine transporter, the intracellular content and mitochondrial membrane potential Potential, Bid, Caspase-3, -8, -9. The changes of cytochrome c in mitochondria and cytoplasm were detected by Western blotting. The experimental results show that WJH-6 has good ability of polyamine transporter recognition and can induce mitochondrial membrane potential reduction, cytochrome c release and Bid, Caspase-3, -8, -9 activation in leukemia K562 and HL-60 cells . The results suggest that WJH-6-induced leukemia K562, HL-60 cell apoptosis and mitochondrial damage.