N-磷酰化二肽甲酯诱导K562细胞凋亡

Apoptosis of K562Cell Induced by N-p hosphoryl Dipeptide Methyl Ester

背景与目的:磷被认为是生命代谢过程中的中心元素,许多N-磷酰化氨基酸和N-磷酰化肽具有重要的生物活性和药用价值,本实验研究N-磷酰化二肽甲酯诱导人慢性髓性白血病K562细胞凋亡作用。方法:采用四甲基偶氮唑盐还原法犤3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl-tetrazoliumbromide,MTT犦检测15种N-磷酰化二肽甲酯对K562细胞的增殖抑制作用;用Hoechst33258染色后观察细胞核的形态学变化;DNA琼脂糖凝胶电泳、双染法和流式细胞术检测细胞凋亡。结果:MTT法筛选出细胞增殖抑制活性较好的化合物是(DIPP-L-Leu)2-L-Lys-OCH3,IC50为22.66μmol/L;Hoechst33258染色后细胞核出现明显的凋亡形态学变化;DNA琼脂糖凝胶电泳出现典型的梯状条带;流式细胞仪检测到早期的凋亡细胞。结论:(DIPP-L-Leu)2-L-Lys-OCH3能够诱导K562细胞凋亡。

Background &Objective:Phosphorus plays a crucial role in me tabolism.Some N-phosphoryl amino a cids and N-phosphopeptides have importa nt biological activities and medicinal value.The current study was designed to investigate the apoptosis of K562cells induced b y N-phosphoryl dipeptide methyl ester.Method:Cell growth inhibition of K562cells induced by 15kinds of N-phosphoryl dipeptide methyl esters was analyzed by using MTT assay.The nuclei were stained by Hoechst 33258and the morphologic ch anges were observed.DNA agarose gel electrophoresis,double-staining,and flow cytomery were used to detect early ap optosis of K562cells.Result:(DIPP-L-Leu)2-L-Lys-OCH 3 was the compound screened with MTT method that had better growt h inhibiting activity with the IC 50 of 22.66μmol /L.Chromatin condensation and nuclear fragmentation were seen under fluorescence microscope in the cells treated with Hoechst 33258.DNA a garose gel electrophoresis showed nuclear fra gmentation(DNA ladder).Early apoptotic cells were also detected by flow cytometry.Conclusion:These results suggest that(DIPP-L-Leu) 2 -L-Lys-OCH 3 could induce the apoptosis of K562cells.