冬凌草活性成分KY3对食管癌EC109细胞增殖的抑制作用

Inhibitory effect of active ingredient of Rabdosia rubescens KY3 on proliferation of esophageal carcinoma EC109 cells

目的:探讨冬凌草活性成分KY3对食管癌细胞EC109增殖的抑制作用及其机制。方法:MTT法测定0、10、20、30、40、60、80、100μmol/L KY3对EC109细胞增殖的影响;流式细胞术检测0、10、20、30μmol/L KY3对EC109细胞凋亡和细胞周期分布的影响,免疫荧光法观察KY3对EC109细胞中微管结构的影响,Hoechst 33258染色观察KY3作用后EC109细胞核的形态。结果:KY3能够抑制EC109细胞的体外增殖;KY3处理后EC109细胞周期阻滞在G2/M期并发生凋亡(P<0.05);KY3作用于EC109细胞后,微管遭到破坏,随着KY3剂量的增加,α-微管蛋白逐渐发生降解并出现细胞核固缩、浓染等典型的凋亡形态。结论:冬凌草活性成分KY3在体外对EC109细胞具有增殖抑制作用,其机制可能与破坏细胞微管结构、G2/M期细胞周期阻滞及诱导细胞凋亡有关。

Aim： To investigate the proliferation inhibition and the mechanism of active ingredients of Rabdosia rubescens KY3 on esophageal carcinoma EC109 cells. Methods： MTT assay was performed to measure the effect of 0,10, 20,30,40,60,80,100 μmol/L KY3 on ECI09 cell proliferation. Flow cytometry assay was conducted to measure the effect of 0,10,20,30 μmol/L KY3 on apoptosis and cell cycle distribution of EC109 ceils, immunofluorescence was used to de- termine the effect of KY3 on microtubule structure of EC109 cells, and Hoechst 33258 staining was used to detect the mor- phological changes of the nucleus of EC109 cells after the treatment of KY3. Results ： KY3 could inhibit the proliferation of EC109 cells in vitro,and induce cell cycle arrest in GJM phase as well as apoptosis of EC109 cells（ P 〈 0.05 ）. After being treated by KY3, the microtubule of EC109 cells was destroyed. After the dose of KY3 increasing, a-microtubulin degradated gradually and had a typical morphological characteristic of apoptosis such as nucleus pycnosis and bright nuclei. Conclusion： The active ingredient of Rabdosia rubescens KY3 could inhibit the proliferation of EC109 ceils in vitro, and the underlying mechanisms may be related to the destruction of microtubule, the cell cycle arrest at GJM phase as well as the induction of the apoptosis of EC109 cells.