EGCG增强食管癌细胞对顺铂化疗敏感性的实验研究

Experimental study on enhancing effect of EGCG,s on chemotherapy sensitivity of Cisplatin in esophageal cancer cell

目的观察表没食子儿茶素没食子酸酯(EGCG)体外增强顺铂(DDP)对食管癌CaEs-17细胞化疗敏感性的作用,并探讨可能的作用机制。方法实验分为空白对照组、DDP组(培养液中DDP终浓度为20μmol/L)、EGCG组(培养液中EGCG终浓度为20 mg/L)、联合组(培养液中EGCG终浓度为20 mg/L,DDP浓度为20μmol/L),各组作用24 h。应用CCK-8法和细胞克隆形成法检测细胞增殖抑制率和存活率,金氏公式评价二药的协同效果;流式细胞仪检测细胞凋亡率及周期分布;Transwell法检测细胞侵袭力;免疫印迹法检测Bcl-2蛋白表达水平。结果 EGCG、DDP均可抑制CaEs-17细胞增殖、诱导细胞凋亡、降低细胞存活率和侵袭力;EGCG可阻滞细胞于G0/G1期,DDP或联用EGCG可阻滞细胞于G2/M期,并能下调Bcl-2蛋白表达水平(P<0.05),二药联用上述作用效果更为显著(P<0.01)。结论 EGCG能够增强DDP对食管癌CaEs-17细胞的化疗敏感性,这种作用部分是通过下调Bcl-2蛋白实现的。

Objective It is to observe the enhancing effect of epigallocatechin -3-gallate （ EGCG） on chemotherapy sen-sitivity of Cisplatin in human esophageal cancer line CaEs -17 in vitro and to explore its possible mechanism .Methods There were four groups：Control group, EGCG group （CaEs-17 cells were pretreated by 20 mg/L EGCG）, Cisplatin group（ CaEs-17 cells were pretreated by 20 mg/L Cisplatin）, EGCG/Cisplatin group （CaEs-17cells were pretreated by 20 mg/L Cisplatin and 20mg/L EGCG）, after treatment for 24 hours, The growth and proliferation of CaEs -17 cell was detected by CCK -8 as-say and Clone formation test and Jin ’ s formula was used to assess the synergistic effects between two drugs .Cell apoptosis and phase were detected by the flow cytometry .Cell invasive ability was measured with Transwell. The expression of Bcl -2 Pro-tein was detected by Western blot .Results EGCG, Cisplatin alone could inhibit cell proliferation , induce cell apoptosis , de-crease cell survival rate and invasive ability and down-regulate the expression of Bcl -2 in CaEs -17 cells （P〈0.05）. EGCG could arrest cell cycle in G 0/G1 phase while Cisplatin alone or combination with EGCG could arrest cell cycle in G 2/M phase.The effect above of Cisplatin combined with EGCG was better than that of Cisplatin or EGCG alone （P〈0.01）.Con-clusion EGCG can enhance chemotherapy sensitivity of Cisplatin on human esophageal cancer Line CaEs -17 in vitro, which is partly related to the down-regulation of Bcl -2 protein on tumor cells .