6-溴异香兰素BVAN08诱发肝癌细胞HepG2纺锤体损伤和有丝分裂灾变死亡  

The Spindle Damage and Mitotic Catastrophe in Hepato-carcinoma HepG2 Cells Induced by Bromovanin BVAN08

作　　者：张博[1] 王林[1] 王豫[1] 徐勤枝[1] 张士猛[1] 周平坤[1] 

机构地区：[1]军事医学科学院放射与辐射医学研究所放射毒理与放射肿瘤学研究室,北京100850

出　　处：《细胞生物学杂志》2008年第5期629-634,共6页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(No.30772592);国家重点基础研究发展规划(973计划)(No.2007CB914603)资助项目~~

摘　　要：研究香兰素衍生物中的6-溴异香兰素(BVAN08)对细胞纺锤体结构的影响及诱发灾变死亡的相关机制,为开发该化合物为新的抗癌药物提供理论依据。通过光学显微镜观察BVAN08作用后细胞形态学变化,流式细胞术检测细胞周期,纺锤体功能检测点实验和原位免疫荧光杂交实验分析细胞有丝分裂进程和纺锤体结构,Western印记检测BVAN08作用后相关蛋白质的变化。结果表明20～60μmol/LBVAN08作用后,HepG2细胞变圆不再贴壁生长、随后脱落死亡,具有浓度依赖性量效关系;明显诱导细胞G2/M期阻滞、导致细胞有丝分裂指数升高,并出现大量的非二倍体和多倍体细胞;破坏细胞纺锤体的结构,多中心体细胞显著增加;该化合物促使细胞周期转录调节因子FoxM1及其下游靶分子细胞周期蛋白B1和Cdk1的降解、阻止有丝分裂过程而导致有丝分裂灾变死亡。研究揭示BVAN08通过破坏纺锤体结构、诱发M期阻滞,导致细胞有丝分裂灾变死亡,FoxM1失活可能参与其作用机制。To investigate the induction of bromovanin （6-bromine-5-hydroxy-4-methoxy-benzaldehyde, BVAN08） on the spindle damage and mitotic catastrophe of cancer cells and its related mechanism, and to provide more solid evidence and experimental data for exploring bromovanin as anticancer new drug. Cellular morphology changes after bromovanin treatment were observed by light microscope, flow cytometry was used to detect cell cycle, spindle checkpoint analysis and immunostaining experiment were performed to investigate the effect of bromovanin on process of mitotic division and spindle structure. Western blot analysis was used to determine protein expression changes. The results demonstrated that HepG2 cells were become rounding and detached quickly after treated with 20-60 umol/L of bomovanin, and at last dead in a dose-dependent manner. Bromovanin treatment resulted in a G2/M arrest, increased mitotic index and considerable ratio of non-diploid or polyploidy cells. Our result also demonstrated the destruction of spindle structure as expressed by multiple centrosome. FoxM1, a cell cycle control associated transcription regulation factor and its downstream target molecules cyclinB1 and Cdkl were down-regulated, and accompanying with the mitotic catastrophe. This study demonstrated that bromovanin destroyed spindle structure, induced mitotic arrest and catastrophe. Inactivation of FoxM1 protein could involve in the effective mechanism of bromovanin.

关 键 词：有丝分裂灾变 纺锤体结构 FOXM1 周期阻滞 

分 类 号：R735.7[医药卫生—肿瘤]