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目的观察姜黄素诱导食管癌Eca-109细胞凋亡,并初步探讨其作用机制。方法通过CCK-8法检测不同浓度、不同时间点作用于Eca-109细胞的增殖抑制率。应用Caspase 3活性检测试剂盒检测胱天蛋白酶3活性。流式细胞技术检测细胞早期凋亡。免疫细胞化学法检测凋亡相关基因Bax和Bcl-2的表达情况。结果随浓度增加时间延长各组生长抑制率有明显增高,且呈剂量浓度依赖性,组间差异有显著性(P<0.01)。胱天蛋白酶3活性检测发现:姜黄素20μmol·L-1,40μmol·L-1作用24 h后OD值与对照组比较,差异具有统计学意义(P<0.05)。流式细胞术检测到姜黄素作用后,细胞凋亡率明显升高。免疫细胞化学染色显示,随着姜黄素浓度的增高和作用时间的增长,Bcl-2蛋白表达减弱,Bax蛋白表达增强。结论姜黄素可能是通过诱导胱天蛋白酶3表达活性增高,上调促凋亡基因Bax及下调抗凋亡基因Bcl-2的表达,诱导Eca-109发生凋亡。
Objective To observe the apoptosis of esophageal carcinoma Eca-109 cells induced by curcumin and to explore its mechanism. Methods The proliferation inhibition rate of Eca-109 cells was detected by CCK-8 assay at different concentrations and at different time points. Caspase 3 activity assay kit was used to detect caspase 3 activity. Flow cytometry to detect cell early apoptosis. Immunocytochemistry was used to detect the expression of apoptosis related genes Bax and Bcl-2. Results With the increase of time, the growth inhibition rate of each group was significantly increased, and in a dose-dependent manner, the difference was significant (P <0.01). The results of caspase 3 assay showed that OD value of curcumin treated with 20μmol·L-1, 40μmol·L-1 for 24 h was significantly lower than that of control group (P <0.05). Flow cytometry detected the role of curcumin, the apoptosis rate was significantly increased. Immunocytochemical staining showed that with the increase of curcumin concentration and action time, the expression of Bcl-2 protein decreased and the expression of Bax protein increased. Conclusion Curcumin induces Eca-109 apoptosis by inducing the increase of caspase 3 expression, up-regulating the expression of Bax and down-regulating the anti-apoptotic gene Bcl-2.