论文部分内容阅读
目的探讨南极土壤来源真菌的次级代谢产物HDN-1对人急性粒细胞白血病NB4细胞的增殖抑制,诱导凋亡作用及其机制。方法采用MTT法检测HDN-1对NB4细胞的增殖抑制作用;DNA结合染料Hoechst 33324染色,Western Blot实验,流式细胞仪检测细胞凋亡以及凋亡相关蛋白的变化。结果 HDN-1能显著抑制NB4细胞的增殖,并呈剂量-时间依赖性;HDN-1作用后,细胞核形态发生明显变化且流式细胞仪分析表明NB4细胞凋亡数量明显增加;HDN-1可引起Caspase-3,8,9激活,bid减少,线粒体中凋亡蛋白Bcl-2,Mcl-1的表达降低,以及γ-H2AX、Parp剪切带增加。但是NB4细胞中融合蛋白PML-RARα的表达不能被HDN-1所抑制。结论 HDN-1诱导凋亡是通过Caspase依赖的线粒体途径和死亡受体途径共同介导的,PML-RARα不是Hsp90的客户蛋白,其诱导凋亡机制可能是通过抑制Hsp90的其他蛋白表达进行的。
Objective To investigate the effects of HDN-1, a secondary metabolite of Antarctic soil-derived fungi, on the proliferation and apoptosis of human acute myeloid leukemia NB4 cells and its mechanism. Methods The inhibitory effect of HDN-1 on the proliferation of NB4 cells was detected by MTT assay. The DNA binding dye Hoechst 33324 staining, Western Blot and flow cytometry were used to detect the changes of apoptosis-related proteins. Results HDN-1 could significantly inhibit the proliferation of NB4 cells in a time-and dose-dependent manner. After HDN-1 treatment, the morphological changes of the cells were observed. Flow cytometry analysis showed that the apoptosis of NB4 cells was significantly increased. Induced the activation of Caspase-3, 8 and 9, decreased the bid, decreased the expression of apoptosis proteins Bcl-2 and Mcl-1 in mitochondria, and increased the γ-H2AX and Parp shear bands. However, the expression of fusion protein PML-RARα in NB4 cells can not be inhibited by HDN-1. Conclusions HDN-1 induced apoptosis is co-mediated by Caspase-dependent mitochondrial pathway and death receptor pathway. PML-RARα is not a Hsp90 client protein. The mechanism of apoptosis induced by HDN-1 may be through inhibition of other protein expression of Hsp90.