摘要
目的体外研究HSP90 C-端抑制剂新生霉素(novobio-cin,NB)与HSP90 N-端抑制剂格尔德霉素(Geldnamycin,GA)的联合应用对Bcr-Abl-的HL-60细胞和Bcr-Abl+的HL-60/Bcr-Abl细胞增殖抑制及诱导凋亡的作用及该作用与细胞色素C释放、Caspase-9/3活性的关系,并且研究序贯应用NB和GA对HL-60细胞的作用。方法细胞用NB和GA联合处理后,采用AO/EB和AnnexinV/PI双染检测细胞凋亡,分光光度法检测Caspase-9/3的活性,用蛋白免疫印迹法检测细胞色素C和procaspase-3的含量。用MTT法检测NB→GA和GA→NB序贯处理对HL-60细胞增殖的抑制作用。结果NB和GA合用抑制HL-60细胞增殖有单独相加的作用;NB和GA合用激活HL-60和HL-60/Bcr-Abl细胞中Caspase-3/9的活性程度高于单独使用NB或GA,协同触发细胞凋亡;预先用NB处理可增强HL-60细胞对GA的敏感性,相比之下,预先用GA处理不影响HL-60细胞对NB的敏感性。结论NB和GA合用可协同抑制白血病细胞生长,诱导细胞凋亡;NB可增强GA对HL-60细胞的抑制能力,而GA不影响NB对HL-60细胞的抑制作用。
Aim To elucidate the effects of cotreatment with HSP90 C-terminal inhibitor NB and HSP90 N-terminal inhibitor GA on proliferation or induction of apoptosis of Bcr-Abl- HL-60 cells and Bcr-Abl ＋ HL-60/ Bcr-Abl cells, and reveal the relationship between apoptosis and mitochondrion pathway; to further investigate the effect of sequence treatment with NB and GA on HL-60 cells. Methods When cells were treated with NB and GA, the typical morphological changes of apoptosis were examined by fluorescence microscopy after AO/EB staining or by flow cytometer with Annexin V/PI staining. The activities of Caspase-9 and Caspase-3 were analyzed by spectrophotometry. The level of cytochrome C in cytosolic and S-100 fraction, and that of procaspase-3 were tested by Western blot. MTT was used to analyze the sequence effects of NB and GA on HL-60 cells. Results Cotreatment with NB and GA exerted additive effect on proliferation inhibition and induction of apoptosis. Cotreatment with NB and GA induced more activation of Caspase-3/9 than treatment with NB or GA alone in HL-60 cells and HL- 60/Bcr-Abl cells. Pretreatment with NB was able to sensitize HL-60 cells to GA;in contrast, pretreatment with GA had no influence on the effect of NB on HL-60 cells. Conclusion Combination of NB and GA had additive effects on proliferation inhibition and apoptosis induction of leukemia cells; NB was able to enhance the effect of GA on HL-60 cells, in contrast, GA had no influence on the effect of NB on HL-60 cells.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2008年第2期165-171,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30171158
30472187)
福建省自然科学基金资助项目(Noco610025)
福建省资助省属高校资助项目(No2005K048)
