摘要
目的研究热休克蛋白90(HSP90)抑制剂17-二甲胺乙胺基-17-去甲氧基格尔德霉素(DMAG)对白血病细胞株HL-60的抑制作用及相关机制。方法通过MTT法、流式细胞术检测HL-60细胞在DMAG作用下增殖及凋亡情况;流式细胞术检测HL-60细胞周期的改变;流式细胞术及Westen blot法检测HL-60细胞Raf蛋白含量;RT-PCR法检测raf-mRNA表达量的变化。结果 DMAG能抑制HL-60细胞的生长,并诱发凋亡。细胞周期分析显示,DMAG作用后,G0/G1期细胞增多,S期细胞减少。DMAG能降低HL-60细胞Raf蛋白的含量,但raf-mRNA表达量无明显变化。结论 DMAG能引起HL-60细胞的凋亡,抑制其生长。DMAG引起HL-60细胞凋亡的机制与降低Raf蛋白含量有关。
Objective To explore the effects of heat shock protein 90（HSP90）-inhibitor,17-dimethylaminoethylamino-17-demethoxygeldanamycin（DMAG）on the leukemia cell line HL-60.Methods The growth curves of HL-60 cells treated with DMAG were made using MTT assay.The apoptosis rate and cell cycle of HL-60 cells treated by DMAG were examined by flow cytometry.The protein level of cytoplasmic Raf was evaluated using FCM and Westen blot,and mRNA level was detected by RT-PCR.Results The proliferation of HL-60 cells was inhibited by DMAG in a dose and time dependent manner,and apoptosis rate increased in the treated cells.Cell cycle analysis showed that the ratio of G0/G1 phase cells significantly increased and the S phase cells ratio decreased after HL-60 cells were treated with DMAG for 48 hours.After the HL-60 cells were treated with DMAG,cytoplasmic protein level of Raf decreased.However,the level of raf-mRNA kept stable.Conclusions DMAG can induce apoptosis and inhibit the growth of HL-60 cells,which may be realized by the depletion of Raf protein.
出处
《临床儿科杂志》
CAS
CSCD
北大核心
2011年第11期1067-1070,共4页
Journal of Clinical Pediatrics
