姜黄素损伤线粒体诱导食管癌Ec-109细胞凋亡的研究

Curcumin induced apoptosis of esophageal cancer cell line Ec-109 cells by damaged mitochondria

目的:探讨姜黄素损伤线粒体诱导食管癌Ec-109细胞凋亡的作用机制。方法:80μmol/mL姜黄素作用于食管癌Ec-109细胞不同时间后,流式细胞仪检测细胞亚二倍体凋亡峰变化及线粒体膜电位变化;蛋白质印迹法检测细胞色素C释放及Caspase-9表达。结果:姜黄素呈时间依赖性诱导食管癌Ec-109细胞凋亡,12、24和48 h细胞亚二倍体凋亡峰分别为(15.89±2.12)%、(26.80±1.87)%和(36.97±1.80)%,对照组为(3.23±0.24)%,总体比较差异有统计学意义,F=6.75,P<0.01,各组间比较差异均有统计学意义,P<0.01。姜黄素作用3、6和12 h后线粒体膜电位分别为84.78%、67.03%和63.16%,对照组为97.17%,差异有统计学意义,F=5.12,P<0.05,各组间比较,6 h组与12 h组差异无统计学意义,P=0.062,余各组间有差别;6 h出现细胞色素C表达,12 h表达最高;12 h检测到Caspase-9表达,24 h表达最高。结论:姜黄素呈时间依赖性诱导食管癌细胞凋亡是通过损伤细胞线粒体、使之释放出细胞色素C以及激活Caspase-9实现的。

OBJECTIVE： To investigate the curcumin induced apoptosis of esophageal cancer cell line Ec-109 cells by damaged mitochondria and explore the mechanism.METHODS：Ec-109 cells were treated with 80 μmol/ml curcumin in different time,and apoptotic Sub-G1 peak and mitochondrial membrane potential were detected by flow cytometry;the expressions of cytochrome C and Caspase-9 were detected by Western-blot.RESULTS：When Ec-109 cells treated with curcumin for 12,24 and 48 h,the apoptotic Sub-G1 peas were about（15.89±2.12）%,（26.80±1.87）% and（36.97±1.80）% respectively.Compared with the control group（（3.23±0.24）%）,there was a statistically significant difference（F=6.75,P〈0.01）.The mitochondrial membrane potential was damaged obviously after 3,6 and 12 h,which was 84.78%,67.03% and 63.16% respectively,and there was a statistically significant difference too（F=5.12,P〈0.05）.And the difference between 6 h and 12 h was not significant（P=0.062）.The expressions of cytochrome C and Caspase-9 appeared a kind of tendency of time-dependent up-regulation after treatment with curcumin for 12 h and 24 h respectively.CONCLUSION：Curcumin can damage mitochondria,from which cytochrome C is released and then Caspase-9 is activated,time-dependently inducuing EC-109 cells apoptosis.