Wnt/β-Catenin信号通路对氧化应激损伤促进食管癌细胞凋亡的调控研究

Oxidative stress injury regulates the apoptosis of esophageal cancer cell by Wnt/β-Catenin signaling pathway

目的:研究H2O2对食管癌细胞凋亡的影响与调控。方法:分别使用1、10、100μmol/L H2O2处理TE-1细胞4 h,构建氧化应激模型;采用台盼蓝拒染方法检测H2O2对食管癌细胞的凋亡作用;利用分光光度计检测H2O2对TE-1细胞丙二醛(MDA)含量和过氧化氢酶(CAT)活力;利用Western Blotting(WB)方法检测氧化应激诱导后凋亡相关蛋白表达变化和对Wnt/β-Catenin信号通路的影响。结果:随着H2O2浓度的增加,TE-1细胞的凋亡率逐步提高;TE-1细胞经H2O2处理后MDA含量提高,CAT酶活力降低,其中100μmol/L组MDA含量最高,为80.77±8.43(P<0.01),100μmol/L组CAT酶活力最低,为16.83±3.96(P<0.01);100μmol/L组与对照组相比,Bcl-2蛋白表达下调,Caspase-3,Bax蛋白表达上调(P<005),Wnt/β-Catenin信号通路相关的Wnt5a、β-Catenin、c-Myc蛋白表达下调。结论:氧化应激可能通过Wnt/β-Catenin信号通路对食管癌TE-1细胞的凋亡产生影响。

Objective:To study the effect and regulation of H2O2 on apoptosis of esophageal cancer cell.Methods:TE-1 cells were treated with 1,10 and 100μmol/L H2O2 for 4 hours respectively to establish oxidativestress model;apoptotic effect of H2O2 on esophageal cancer cell was detected by trypan blue staining method;malondialdehyde(MDA)content and catalase(CAT)activity of H2O2 on TE-1 cells were detected byspectrophotometer;Western Blotting(WB)was used to detect the expression of apoptosis-related proteinsinduced by oxidative stress and its effect on Wnt/β-Catenin signaling pathway.Results:As the concentration ofH2O2 increases,the apoptosis rate of TE-1 cells gradually increases;The content of MDA in TE-1 cells wasincreased after H2O2 treatment,and the activity of CAT was decreased.The content of MDA in 100μmol/L groupwas the highest,which was 80.77±8.43(P<0.01).The activity of CAT in 100μmol/L group was the lowest,being 16.83±3.96(P<0.01);Compared with the control group,Bcl-2 protein expression was down-regulated,Caspase-3 and Bax protein expression were up-regulated in the 100μmol/L group(P<0.05),Wnt/β-Cateninsignaling pathway-related Wnt5 a,β-Catenin,c-Myc protein The expression is down-regulated.Conclusion:Oxidative stress may affect the apoptosis of esophageal cancer TE-1 cells through Wnt/β-Catenin signalingpathway.