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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。
Objective To study the effect of curcumin on the cell cycle of human hepatocellular carcinoma HepG2 and the role of microtubule system in it. Methods The inhibitory effect of curcumin on HepG2 cell proliferation was detected by MTT assay. The effect of curcumin on the cell cycle distribution of HepG2 cells was analyzed by flow cytometry. The effect of curcumin on the microtubule structure of HepG2 cells was observed by laser scanning confocal microscopy. The effect of curcumin on the expression of tubulin α-tubulin was examined. In vitro, the effect of curcumin on tubulin polymerization and depolymerization activity was observed. Results Curcumin had an inhibitory effect on the proliferation of HepG2 cells and was related to time and dose. With the increase of curcumin concentration, the proportion of HepG2 cells arrested in G2 / M phase also increased gradually. The results of laser scanning confocal microscopy showed that curcumin was significantly Destroy the microtubule structure and change the state of tubulin polymerization. Western blotting showed that the decrease of α-tubulin protein expression was related to the drug concentration. Curcumin can also affect tubulin polymerization and depolymerization activity. Conclusion Curcumin inhibits the growth of HepG2 cells by disrupting the microtubule structure and down-regulating the expression of tubulin α-tubulin in HepG2 cells, and arresting HepG2 cells in G2 / M phase.