雷公藤内酯醇联合顺铂对食管鳞癌细胞ECA-109增殖的影响

Effect of triptolide combined with cisplatin on proliferation of esophageal cancer ECA-109 cells

目的观察雷公藤内酯醇(TPL)以及TPL联合顺铂对人食管鳞癌细胞ECA-109增殖的影响,并探讨其作用机制。方法选取对数生长期的人ECA-109细胞,随机分为空白对照组(不经药物处理)、不同浓度(6.25、12.50、25.00、50.00、100.00μg·L^(-1))TPL组、4.17μmol·L^(-1)顺铂组、TPL(25.00μg·L^(-1))联合顺铂(4.17μmol·L^(-1))组;采用四甲基偶氮唑蓝法测定各组不同时间段(24、48、72 h)的细胞增殖抑制率,并计算TPL与顺铂之间的交互作用;采用Western blot检测空白对照组、25.00μg·L^(-1)TPL处理组、4.17μmol·L^(-1)顺铂组、TPL联合顺铂组细胞培养24 h后半胱氨酸天冬氨酸特异性蛋白酶-3(caspase-3)的表达。结果同一时间点,不同浓度TPL组对ECA-109细胞增殖的抑制率显著高于空白对照组(P<0.05),且呈剂量依赖性(P<0.05);同一药物浓度条件下,随作用时间延长,细胞增殖抑制率显著增加(P<0.05),即呈时间依赖性。同一时间点,TPL联合顺铂组对ECA-109的抑制率高于同药物浓度条件下单用顺铂或单用TPL对细胞增殖的抑制率(P<0.05)。加药培养24 h后,TPL组、顺铂组及TPL联合顺铂组caspase-3蛋白表达量均显著高于空白对照组(P<0.05),且TPL联合顺铂组caspase-3蛋白表达量高于同药物浓度条件下单用TPL组、单用顺铂组蛋白表达量(P<0.05)。结论 TPL具有抑制食管癌ECA-109细胞增殖的作用,TPL联合顺铂有协同作用,增加对食管癌ECA-109细胞的抑制,其机制可能与促进caspase-3蛋白表达有关。

Objective To observe the effect of triptolide combined with cisplatin on proliferation of esophageal cancer ECA-109 cells and to explore its potential mechanisms. Methods ECA-109 cells in exponential phase of growth were randomly divided into blank control group,triptolide group（ 6. 25,12. 50,25. 00,50. 00,100. 00 μg·L^-1）,4. 17 μmol·L^-1 cisplatin group and TPL ＋ cisplatin group（ 25. 00 μg · L^-1 triptolide ＋ 4. 17 μmol · L^-1 cisplatin）. Methylthiazolyl tetrazolium method was used to evaluate the cell proliferation inhibition rate in each group at different time（ 24,48,72 h） and the interaction between TPL and cisplatin was calculated. Western blot was used to detect the expression of cysteinyl aspartate specific proteinase-3（ caspase-3） in blank control group,25. 00 μg · L^-1 TPL group,cisplatin group and TPL ＋ cisplatin group after being cultured for 24 hours. Results The proliferation inhibition rates were significantly higher in the different concentration triptolide groups than those in blank control group at the same time point（ P〈0. 05）,and the proliferation inhibition rate in triptolide group was concentration dependent（ P〈0. 05）; at the same drug concentration,the proliferation inhibition rates in the different concentration triptolide groups were increased with the time,it showed time dependent（ P〈0. 05）. At the same time point,the proliferation inhibition rate in TPL ＋ cisplatin group was significantly higher than that in different concentration triptolide group and cisplatin group（ P〈0. 05）. After being cultured for 24 h,the expression of caspase-3 in triptolide group,cisplatin group and TPL ＋ cisplatin group was obviously higher than that in the blank control group. Besides,the expression of caspase-3 in TPL ＋ cisplatin group was significantly higher than that in triptolide group and cisplatin group（ P〈0. 05）.Conclusion Triptolide,as well as combined with cisplatin,can inhibit the proliferation of esophageal cancer ECA-109 cells,and its potential mechanism might be involved in promoting the expression of caspase-3 protein.