[关键词]
[摘要]
目的 研究姜黄素对人肝癌HepG2细胞周期的影响及微管系统在其中的作用。方法 MTT法检测姜黄素对人肝癌HepG2细胞增殖的抑制作用;流式细胞仪分析姜黄素对HepG2细胞周期分布的影响;激光共聚焦显微镜观察姜黄素对HepG2细胞微管结构变化的影响;Western blotting 检测姜黄素对微管蛋白α-tubulin表达的影响;在离体条件下,观察姜黄素对微管蛋白聚合和解聚活性的影响。结果 姜黄素对HepG2细胞增殖具有抑制作用且与时间和剂量相关;随着姜黄素浓度的升高,HepG2细胞阻滞于G2/M期的比例也逐渐增加;激光共聚焦显微镜观察可见姜黄素明显破坏细胞微管结构,改变细胞微管蛋白聚合状态的发生;Western blotting检测发现细胞内α-tubulin蛋白表达的减弱与试药浓度相关。姜黄素还可影响微管蛋白聚合和解聚活性。结论 姜黄素通过破坏HepG2细胞的微管结构及下调微管蛋白α-tubulin的表达,将HepG2细胞阻滞于G2/M期,从而抑制肝癌HepG2细胞的生长。
[Key word]
[Abstract]
Objective To investigate the effect of curcumin (Cur) on cycle in HepG2 cells and the action of microtubule system for it. Methods The proliferation inhibition of Cur on HepG2 cells was evaluated by MTT assay; The effect of Cur on the cycle of HepG2 cells was analyzed by flow cytometry; Confocal laser scanning microscopy (CLSM) was used to observe the changes of microtubule in HepG2 cells after treated with Cur; Western blotting was performed to determine the expression of α-tubulin; The in vitro effects of Cur on tubulin polymerization and depolymerization activities were studied. Results Cur inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner. With Cur concentration increasing, the proportion of HepG2 cells arrested in G2/M phase was also increased. CLSM showed that Cur could seriously destroy the cell microtubule structure and change the polymerization state of tubulin. Western blotting showed that the decrease of the expression level of α-tubulin was related to Cur concentration. Cur could also interfere the microtubule polymerization and depolymerization activities. Conclusion Cur could inhibit the proliferation of HepG2 cells by destroying the microtubule structure and downregulating the expression of α-tubulin, and make HepG2 cells arrested in G2/M phase.
[中图分类号]
[基金项目]
国家重点基础研究发展计划(“973”计划)资助项目(2010CB833603);海外及港澳学者合作研究基金资助项目(31129002);暨南大学科研培育与创新基金研究项目(21612601)