乙酰唑胺对食管癌细胞系E9706的放射增敏作用

The radiosensitive effect of acetazolammide on esophageal carcinoma cell lines E9706

目的研究碳酸酐酶IX抑制剂乙酰唑胺对食管癌细胞系E9706的放射增敏作用及相关机制。方法 MTT实验确定1μM浓度乙酰唑胺为放射增敏研究中的药物浓度。1μM乙酰唑胺处理E9706细胞24h后,给以不同剂量(0、2、4、6、8Gy)6MV-X线照射,克隆形成实验检测乙酰唑胺放射增敏作用。流式细胞仪进行细胞周期分析,蛋白印迹实验检测凋亡相关蛋白的表达情况。结果乙酰唑胺对E9706细胞有浓度时间依赖性抑制作用,1μM乙酰唑胺有放射增敏作用,增敏比为1.40。细胞周期分析显示乙酰唑胺增加E9706细胞G2/M期细胞比例。蛋白印迹实验发现乙酰唑胺上调Bax蛋白表达,下调Bcl-2蛋白表达。结论碳酸酐酶IX抑制剂乙酰唑胺在食管癌细胞系E9706中有放射增敏作用,增加G2/M期细胞比例及诱导凋亡参与了其放射增敏作用。

Objective To investigate the radiosensitizing effect and mechanisms of acetazolammide,a carbonic anhydrase IX inhibitor,on esophageal carcinoma cell lines E9706.Methods Inhibitory effect of acetazolamide on E9706 cells was assayed by MTT assay.After incubation with 1μM acetazolammide,E9706 cells were irradiated by X-ray at different doses（0,2,4,6,8 Gy）.The cells were assayed for clonogenic survival to determine the radiosensitizing effect of acetazolammide.Flow cytometry was used to analyze cell cycle.Western blot was used to detect the expression of Bax and Bcl-2.Results Acetazolamide had a dose-dependent and time-dependent inhibitory effect on E9706 cells.At the concentration of 1μM,acetazolamide has radiosensitizing effect on E9706 and the sensitivity enhancement ratio（SER）was 1.40.Cell cycle analysis showed that acetazolamide increased the proportion of cells in G2/M phase.Western blot showed that acetazolamide upregulated the expression of Bax protein,down regulated the expression of Bcl-2 protein.Conclusion Acetazolamide had radiosensitizing effect on esophageal cancer cell lines E9706 by increasing the proportion of cells in G2/M phase and induced apoptosis.