EGCG对5-氟尿嘧啶用于食管癌CaEs-17细胞增加疗效的实验研究

Effect of EGCG's in enhancing chemotherapy sensitivity of 5-fluorouracil on human esophageal cancer line CaEs-17 in vitro

目的观察表没食子儿茶素没食子酸酯(EGCG)体外增强5-氟尿嘧啶(5-Fu)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、5-Fu组(培养液中5-Fu终浓度为6μmol/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,5-Fu浓度为6μmol/L),4组作用24 h,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;免疫印迹法检测突变型P53(mtP53)蛋白表达水平。结果 EGCG、5-Fu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、5-Fu均可阻滞细胞于G0/G1期,并能下调mtP53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5-Fu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mtP53蛋白实现的。

Objective To observe the effect of epigallocatechin-3-gallate （ EGCG ） in enhancing chemotherapy sensitivity of 5-fluorouracil （ 5-Fu ） on human esophageal cancer line-CaEs-17 in vitro, and to explore possible action mechanism.Methods The experiment consisted of four groups：blank control group,5-Fu group （ final concentration of 5-Fu：6μmol/L in culture medium）,EGCG group （final concentration of EGCG：20mg/L）,combination group （final concentration of EGCG：20mg/L and 5-Fu：6μmol/L）,affected for 24 hours in four groups.The inhibitory rate on cell proliferation and survival rate were detected by CCK-8 method and cell clone formation test,respevtively;the synergism effect of two drugs was evaluated according to Jin’ s formula;the cell apoptosis rate and cell cycle distribution were detected by flow cytometry;cell invasiveness was determined by Transwell method;the expression levels of mutant P53 （mtP53） protein were detected by Western Blot. Results Both EGCG and 5-Fu could inhibit the proliferation of CaEs-17 cells,induce cell apoptosis ,decrease cell survival rate and cell invasiveness, moreover, both EGCG and 5-Fu could block cell at G0/G1 stage and cloud down-regulate the expression levels of mtP53 protein （ P 〈0.05）,the effects of combined application of two drugs were more obvious （ P 〈0.01）.Conclusion EGCG can enhance chemotherapy sensitivity of 5-Fu on human esophageal cancer line-CaEs-17 in vitro, which is performed partly through down-regulating the expression levels of mutation P53 protein.