白藜芦醇激活结肠上皮细胞存活途径及结肠癌细胞凋亡信号通路

Resveratrol activates survival pathway in colon epithelial cells and apoptotic signaling pathway in colon cancer cells

目的研究白藜芦醇(RES)对结肠上皮NCM460细胞和结肠癌HCT116细胞生长、凋亡及MAPK信号通路的影响。方法体外培养NCM460和HCT116细胞,采用0、10、20、40μM的RES处理细胞72 h,倒置显微镜观察细胞形态及克隆数,流式细胞术分析细胞周期和凋亡率,Western blot检测Erk1/2、p-Erk1/2、P38 MAPK、p-P38 MAPK、JNK1/2、p-JNK1/2和c-Myc蛋白表达水平。结果RES对HCT116细胞形态和克隆的抑制作用均较NCM460细胞更显著(P<0.05);RES(20和40μM)处理HCT116细胞后,G 1期细胞分布减少,细胞凋亡率升高(P<0.05),而对NCM460细胞的作用相对较弱些;另外,RES上调NCM460细胞中ERK1/2磷酸化水平(P<0.05),而对HCT116细胞中ERK1/2磷酸化没有影响;RES诱导HCT116细胞中JNK磷酸化水平,抑制c-Myc蛋白表达(P<0.05),而对NCM460细胞中p-JNK和c-Myc蛋白表达水平没有影响。结论RES通过诱导HCT116细胞中凋亡信号的活化发挥抗肿瘤作用,而通过激活NCM460细胞中存活信号降低药物毒性。

Objective To study the impacts of resveratrol(RES)on the growth,apoptosis and MAPK signaling pathway in colon epithelial cells(NCM460)and colon cancer cells(HCT116).Methods NCM460 and HCT116 cells were cultured in vitro and treated with RES(0,10,20,and 40μM)for 72 h.The morphology and colonies of NCM460 and HCT116 cells were detected using inverted microscope.Flow cytometry was used to measure the cell cycle and apoptotic rate.The protein expression levels of extracellular-regulated kinase 1/2(Erk1/2),p-Erk1/2,mitogen-activated protein kinase(P38 MAPK),p-P38 MAPK,JNK1/2,p-JNK1 and ellular myelocytomatosis oncogene(c-Myc)were determined by Western blot.Results The inhibitory effect of RES on the morphology and colonies in HCT116 cells was more significant than NCM460 cells(P<0.05).RES exposure(20 and 40μM)led to the reduction of G 1 phase cell distributio and an induction of apoptosis in HCT116 cells(P<0.05),but to a much lesser extent in NCM460 cells.Furthermore,RES upregulated the phosphorylation of ERK1/2 in NCM460 cells but not in HCT116 cells(P<0.05).In contrast,RES enhanced the phosphorylation of JNK and decreased c-Myc expression in HCT116 cells but not in NCM460 cells(P<0.05).Conclusion RES exerted its antitumor effect by inducing activation of apoptotic signaling pathway in HCT116 cells,while reducing drug toxicity by activating survival pathway in NCM460 cells.