摘要
目的:探讨c-Jun氨基末端激酶(JNK)信号通路在D-氨基葡萄糖衍生物2-(3-羧基-1-丙酰氨基)-2-脱氧-D-葡萄糖(COPADG)诱导人食管癌Eca-109细胞凋亡中的作用及机制.方法:体外培养Eca-109细胞,用COPADG及特异性JNK抑制剂SP600125对Eca-109细胞进行处理;Western印迹法检测P-JNK蛋白表达的变化;MTT法检测不同时间点细胞增殖抑制率;倒置相差显微镜下观察细胞形态学变化,AnnexinV/PI染色结合流式细胞术检测细胞凋亡率.结果:Western印迹显示随着COPADG作用浓度增加P-JNK蛋白表达逐渐增强,经SP600125预处理后,COPADG诱导Eca-109细胞P-JNK蛋白表达明显减弱(P<0.01),COPADG诱导的细胞凋亡率明显减低,细胞增殖抑制率显著下降,与COPADG单作用组之间比较差异有统计学意义(P<0.01).结论:JNK信号转导通路在COPADG诱导Eca-109细胞凋亡过程中发挥重要作用,COPADG通过JNK信号通路诱导Eca-109细胞凋亡.
AIM： To investigate the role of c-jun N-terminal kinase （JNK） signaling pathway in the apoptosis of human esophageal cancer Eca-109 cells induced by the derivative of D-glueosamine （ { 2-（ 3-carboxy-1-oxopropyl ） amino-2-deoxy-D-Glucose } , COPADG） and the possible mechanisms. METHODS ： Eca-109 cells were cultured in vitro by using RPMI-1640 and calf serum. Eca-109 cells were pre-incubated with SP600125 for 30 min prior to exposure to COPADG at different concentrations and for different time. Changes in expression of P-JNK protein were examined by Western Blot; cell growth inhibitory rate was detected by MTT colorimetric assay; the cell morphological changes were observed by inverted phase contrast microscopy. Apoptosis rate was analyzed by using Annexin V/PI fluorescence staining together with flow cytometry. RESULTS： COPADG could significantly inhibit the proliferation of Eca-109 cells and induce their apoptosis. Western Blot showed that the protein expression of P-JNK was increased in a dose-dependent manner in Eca-109 cells after stimulated by COPADG. SP600125 remarkablely decreased the protein expression of P-JNK as well as the apoptosis rate and cell growth inhibitory rate in Eca-109 cells induced by COPADG as compared with those treated with only COPADG. CONCLUSION： JNK signaling pathway may play an important role in the apoptosis of Eca-109 cells following COPADG treatment.
出处
《第四军医大学学报》
北大核心
2007年第7期616-618,共3页
Journal of the Fourth Military Medical University
基金
中科院西部之光项目(200524号)
