NF-κB信号通路在七方胃痛颗粒调控H.pylori感染人胃腺癌细胞TFF1表达中的作用  

QIFANGWEITONG GRANULES THROUGH NF-κB SIGNAL PATHWAY UP-REGULATES TFF1 EXPRESSION AFTER HELICOBACTER PYLORI INFECTED IN HUMAN GASTRIC CANCER CELLS IN VITRO

作　　者：曾光[1] 陈国忠[1] 莫喜晶[1] 夏李明[1] 赵学尧[1] 廖纬琳[1] 邓庆圆[1] 

机构地区：[1]广西中医药大学第一附属医院消化内科,南宁530023

出　　处：《广西医科大学学报》2012年第6期837-840,共4页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.30960474);广西科技开发项目(No.10183011);广西教育厅科研项目(No.2007-153)

摘　　要：目的:探讨核因子-κB(nuclear factor kappa B,NF-κB)信号通路在七方胃痛颗粒对幽门螺杆菌(Helicobacter pylori,H.pylori)感染人胃腺癌细胞(human gastric cancer cells,AGS)三叶肽因子1(trefoil factor family 1,TFF1)表达中的作用。方法:采用实时荧光定量PCR观察H.pylori感染AGS TFF1mRNA的表达;双荧光报告基因系统检测质粒载体TFF1(Wt)野生株和多个变异株报告基因瞬时转染AGS,TFF1基因转录情况及七方胃痛颗粒的干预效果,并分析TFF1启动子区NF-κB蛋白结合位点碱基序列区间;凝胶阻滞电泳(EMSA)确定TFF1启动子区NF-κB蛋白结合位点和七方胃痛颗粒对其的作用。结果:药物血清组TFF1mRNA表达及TFF1表达与其它组比较差异有统计学意义(P<0.05),TFF1(Wt)野生株和变异株基因转录表达较空白组均显著增加(P<0.01),TFF1(A)变异株基因表达显著低于Wt株(P<0.01),和其他TFF1变异株相比,差异有统计学意义(P<0.05),变异株之间比较差异无统计学意义(P>0.05);七方胃痛颗粒可降低NF-κB的DNA活性。结论:七方胃痛颗粒上调H.pylori感染AGS TFF1的表达,可能是通过NF-κB信号通路实现的。Objective： To study the effect of Qifangweitong granules on trefoil factor iamlly 1 （TFF1） expression through nuclear factor kappa B （NF-κB） signal pathway after Helieobacter pylori （H. pylori） infection in human gastric cancer cells （AGS） in vitro. Methods： Fluorescence quantitative PCR was applied to detect the TFF1 mRNA expression after H. pylori infected AGS. Based on reporter gene of plasmid vector TFF1 wild strain and several variant strains transient transfection in AGS, gene transcription of TFF1 and intervention effect of Qifangweitong granules were observed, and base sequence of NF-κB protein bining site on TFF1 gene promoter region was determined. Electrophoresis mobility shift assay （EMSA） was used to define NF-κB protein binding site on TFF1 gene promoter region and the effect of the Qifangweitong granules on it. Results： TFF1 mRNA and TFF1 gene expression of drug serum group was statistically significant differences between drug serum group and other groups （ P〈0.05）. After transfection, TFF1 Wt gene and mutant genes transcription were all significantly increased than unstimulated cells after H. pylori stimulation （ P〈0.01）. But TFF1 A mutant gene is significantly lower than Wt gene （ P〈0. 01） and other mutant TFF1 genes （ P〈0. 05）. There was no significant difference between mutant TFF1 genes （ P〈0.05）. Activity of NF-κB binding DNA could be decreased by Qifangweitong granules. Conclusion： Qifangweitong granules can promote TFF1 expression after AGS infected by H. pylori, and it may be related to the NF-κB signal pathway.

关 键 词：NF-ΚB通路 七方胃痛颗粒 人胃腺癌细胞 三叶肽因子1 瞬时转染 凝胶阻滞电泳 

分 类 号：R735.2[医药卫生—肿瘤]