新生霉素抑制HL-60／Bcr-Abl细胞增殖与阻断Hsp90伴侣功能的关系  

Inhibition of novobiocin on proliferation of HL-60/Bcr-Abl cells involves disruption of Hsp90 chaperon function

作　　者：吴丽贤[1] 许建华[1] 张昆仲[1] 温彩霞[1] 黄秀旺[1] 

机构地区：[1]福建医科大学药理系,福建医科大学临床药理研究所,福建福州350004

出　　处：《中国药理学通报》2008年第1期14-19,共6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(NoC0610025);福建省属高校资助项目(No2005k048)

摘　　要：目的研究新生霉素(novobiocin,NB)对HL-60/Bcr-Abl细胞的作用,并探讨该作用是否与Bcr-Abl水平和热休克蛋白90(Hsp90)分子伴侣的功能有关。方法用蛋白免疫印迹法检测Bcr-Abl的含量,采用免疫共沉淀的方法研究Hsp90分子伴侣的功能。先将Bcr-Abl与其分子伴侣共沉淀下来,然后用免疫印迹法检测沉淀物中与Bcr-Abl结合的Hsp90和Hsp70含量变化。应用蛋白酶体抑制剂ALLnL研究Bcr-Abl降解的途径。结果NB能降低HL-60/Bcr-Abl细胞中Bcr-Abl的蛋白水平,NB使Bcr-Abl与Hsp90和Hsp70的结合减少,促进蛋白酶体降解Bcr-Abl蛋白,ALLnL处理后NP-40insoluble部分Bcr-Abl蛋白水平提高。结论该研究证明NB通过干扰Hsp90伴侣功能,减少Bcr-Abl与Hsp90和辅伴侣的结合,从而促进蛋白酶体降解Bcr-Abl蛋白,最终抑制HL-60/Bcr-Abl细胞增殖。Aim In an effort to confirm the effects of NB on Bcr-Abl expressing HL-60/Bcr-Abl cells, and the relationship between these effects and the molecular chaperone functions of heat shock protein 90 （Hsp90）. Methods The amounts of Bcr-Abl protein were tested by Western blot. Molecular chaperone functions of Hsp90 were measured by coimmunoprecipitation. Co-immunoprecipitation of Bcr-Abl and its molecular chaperones, the immunoprecipitate was then subjected to Western blot analysis with anti-Abl, antiHsp90, or anti-Hsp70 mAb. Proteasome inhibitor ALLnL was used to determine the down-regulation pathway of Bcr-Abl. Results An exposure of HL-60/ Bcr-Abl cells to NB produced down-regulation of intra-cellular Bcr-Abl protein levels. By binding and inhibi- ting Hsp90, NB treatment decreased the binding of Bcr-Abl with Hsp90 and Hsp70 and induced the proteasomal degradation of Bcr-Abl, because cotreatment with proteasome inhibitor ALLnL increased Bcr-Abl levels in NP40-insoluble fraction. Conclusions These studies demonstrate for the first time the activity of NB against HL-60/Bcr-Abl cells and suggest that destruction of Hsp90 chaperon function, resulting in disruption the combination of Bcr-Abl with Hsp90 and cochaperons, may be an important mechanism of NB-media- ted effects on HL-60/Bcr-Abl cells.

关 键 词：新生霉素 Bcr-Abl蛋白 热休克蛋白90 分子伴侣 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]