一种N取代靛红衍生物对肝癌细胞的作用及机制

Effect and mechanism of an N-substituted isatin derivative on hepatocellular carcinoma cells

目的探讨一种新型N取代靛红衍生物(3MMIS)诱导肝癌细胞Hep G2产生周期阻滞并死亡的相关机制。方法利用酸性磷酸酶法检测体外抗癌活性。光镜下观察细胞的形态变化特征。流式细胞计检测细胞周期。Western blot检测细胞周期相关蛋白水平。检测3MMIS对微管组装的影响。结果 3MMIS对Hep G2细胞的IC50为0.4 mg/L。经过药物处理后细胞变圆,并出现很多囊泡。药物作用后细胞周期被阻滞在G2/M期,Cyclin B1持续增多,cyclin D1、cyclin E、E2F1、Rb和c-Myc的蛋白水平显著下调。3MMIS作用下细胞内可溶性游离微管蛋白增多,不溶性聚合微管蛋白减少。结论 3MMIS可以通过干扰微管蛋白的聚合引起Hep G2细胞发生周期阻滞,从而诱导细胞死亡。

Objective To investigate the mechanism of a novel N-substituted isatin derivative（ 3 MMIS） that can induce the cell cycle arrest and death of Hep G2 cells. Methods Antitumor activity was detected by acid phosphatase assay. The morphological changes of cells were observed by microscopy at different time intervals. Flow cytometry was used to detect the cell cycle. Western blot was used to detect the effect of 3 MMIS on cell cycle-related proteins. The effect of 3 MMIS on microtubule assembly was detected. Results The IC50 of 3 MMIS-treaded Hep G2 cells was 0. 4 mg/L. Light microscopy showed that the cells exposed to the drug became rounded up,and developed a lot of vesicles in the cells. Flow cytometry showed that the cell cycle was blocked in G2/M phase after drug treatment. Western blot showed abnormal expression of cycle related proteins.3 MMIS treatment increased soluble and free tubulins but decreased insoluble and polymenized tubulins in Hep G2 cells. Conclusions 3 MMIS can induce cell death in Hep G2 cells by interfering the polymerization of tubulins and casing cell cycle arrest at the G2/M phase.