藤黄酸对人结肠癌HT-29细胞生物学行为及JNK信号通路的影响

目的探讨藤黄酸对人结肠癌HT-29细胞增殖、凋亡、迁移及ASK-1、p-ASK-1、JNK、p-JNK表达的影响。方法将人结肠癌HT-29细胞分为空白对照组与藤黄酸组(0.5、1.0、2.5、5.0μmol/ml)。不同浓度藤黄酸干预24、48、72h后,MTT方法检测藤黄酸对HT-29细胞增殖的抑制作用。2.5μmol/ml藤黄酸干预48 h后,流式细胞术检测HT-29细胞的凋亡率;划痕实验检测藤黄酸对HT-29细胞迁移能力的影响;Western blot实验检测藤黄酸对HT-29细胞JNK、p-JNK、ASK-1、p-ASK-1的表达。结果不同浓度藤黄酸均能显著抑制HT-29细胞增殖,且呈时间和浓度依赖性。2.5μmol/ml藤黄酸干预48 h后,HT-29细胞凋亡率升高,迁移能力降低(P<0.01);JNK、p-JNK、ASK-1、p-ASK-1蛋白表达水平均显著升高(P<0.01)。结论藤黄酸能够抑制人结肠癌HT-29细胞增殖与迁移,并促进凋亡,其作用机制可能与调控JNK信号通路相关。

Effects of gambogic acid on biological behavior and JNK signaling pathway of human colon cancer HT-29 cells

Objective To investigate the effects of gambogic acid on the proliferation,apoptosis,migration and the expression of ASK-1、p-ASK-1、JNK、p-JNK in human colon cancer HT-29 cells.Methods HT-29 cells were divided into blank control group and gambogic acid group(0.5,1.0,2.5,5.0μmol/ml).The inhibitory effect of gambogic acid on HT-29 cell proliferation was detected by MTT method after 24,48 and 72 hours of intervention with different concentrations of gambogic acid.After 48 hours of intervention with 2.5μmol/ml gambogic acid,the apoptosis rate of HT-29 cells was detected by flow cytometry;the effect of gambogic acid on the migration ability of HT-29 cells was detected by scratch test;the expression of ASK-1,p-ASK-1,JNK,p-JNK by Western blot method.Results Different concentrations of gambogic acid could significantly inhibit the proliferation of HT-29 cells in a time and concentration dependent manner.After 48 hours of intervention with 2.5μmol/ml gambogic acid,the apoptosis rate of HT-29 cells was increased,migration ability of HT-29 cells was decreased(P<0.01),and the expression of ASK-1、p-ASK-1、JNK、p-JNK was increased significantly(P<0.01).Conclusion Gambogic acid could inhibit the proliferation and migration of HT-29 cells,and promote apoptosis,and its mechanism might be related to the regulation of JNK signaling pathway.