幽门螺杆菌感染对胃癌细胞株恶性转化的影响及机制研究  

Effect of Helicobacter pylori infection on malignant transformation of gastric cancer cell lines and its mechanism

作　　者：吕汪霞[1] 方阳 

机构地区：[1]浙江省肿瘤医院腹部肿瘤内科,浙江杭州310022 [2]温州医科大学医学与生命科学学院,浙江温州325035

出　　处：《中华全科医学》2018年第3期383-387,共5页Chinese Journal of General Practice

基　　金：浙江省自然科学基金(LY14H160011)

摘　　要：目的探讨幽门螺杆菌感染对胃癌细胞株恶性转化的影响并探讨其可能的机制。方法将胃癌AGS细胞分为细胞毒素相关基因A(Cag A)质粒组(转染p CDNA3.1-Cag A质粒)和阴性对照组(转染p CDNA3.1空白质粒)以及Cag A质粒组(转染p CDNA3.1-Cag A质粒和p EGFP-C1空白质粒)、Cag A+程序化细胞死亡因子4(PDCD4)质粒组(转染p CDNA3.1-Cag A质粒和p EGFP-C1-PDCD4质粒)和阴性对照组(转染p CDNA3.1空白质粒和p EGFP-C1空白质粒)。采用逆转录-聚合酶链反应(RT-PCR)进行PDCD4、Twist1、上皮细胞钙粘蛋白E(E-cadherin)、波形蛋白(Vimentin)mRNA水平,采用蛋白质印迹(Western blot)法测定各组细胞PDCD4、Twist1、E-cadherin、Vimentin蛋白水平,采用transwell小室测定细胞侵袭能力。结果 Cag A质粒组Twist1 mRNA、Vimentin mRNA表达量高于阴性对照组(P<0.05),PDCD4 mRNA、E-cadherin mRNA表达量低于阴性对照组(P<0.05);Cag A+PDCD4质粒组Twist1 mRNA、Vimentin mRNA表达量低于Cag A质粒组(P<0.05),PDCD4 mRNA、E-cadherin mRNA表达量高于Cag A质粒组(P<0.05)。Cag A质粒组Twist1蛋白、Vimentin蛋白表达量高于阴性对照组(P<0.05),PDCD4蛋白、E-cadherin蛋白表达量低于阴性对照组(P<0.05);Cag A+PDCD4质粒组Twist1蛋白、Vimentin蛋白表达量低于Cag A质粒组(P<0.05),PDCD4蛋白、E-cadherin蛋白表达量高于Cag A质粒组(P<0.05)。Cag A质粒组侵袭细胞数高于阴性对照组(P<0.05),Cag A+PDCD4质粒组侵袭细胞数低于Cag A质粒组(P<0.05)。结论幽门螺杆菌感染可增加胃癌细胞的侵袭能力,其机制可能为幽门螺杆菌独立因子Cag A通过抑制PDCD4表达促进胃癌细胞的上皮-间质转化。Objective To investigate the effect of Helicobacter pylori infection on the malignant transformation of gastric cancer cell lines and to explore its possible mechanism. Methods The gastric cancer AGS cells were divided into cytotoxin related gene A（ Cag A） plasmid group（ transfected with p CDNA3. 1-Cag A plasmid） and negative control group（ transfected with p CDNA3. 1 blank plasmid）,and were divided into Cag A plasmid group（ transfected with PCDNA3. 1-Cag A plasmid and p EGFP-C1-PDCD4 plasmid）,the Cag A ＋ programmed cell death factor 4（ PDCD4） plasmid group（ transfected with p CDNA3. 1-Cag A plasmid and p EGFP-C1-PDCD4 plasmid） and Negative control group（ transfected with p CDNA3. 1 blank plasmid and p EGFP-C1 blank plasmid）. The levels of PDCD4,Twist1,E-cadherin,Vimentin mRNA in cells were measured by reverse transcription-polymerase chain reaction（ RT-PCR）. The protein levels of PDCD4,Twist1,E-cadherin and Vimentin were determined by Western blot,cell invasion was measured using transwell chamber. Results The expression of Twist1 mRNA and Vimentin mRNA in Cag A plasmid group were higher than those in negative control group（ P〈 0. 05）. The expression of PDCD4 mRNA and E-cadherin mRNA in Cag A plasmid group were lower than those of negative control group（ P〈 0. 05）. The expression of Twist1 mRNA and Vimentin mRNA in Cag A ＋ PDCD4 plasmid group were lower than those in Cag A plasmid group（ P〈 0. 05）. The expression of PDCD4 mRNA and E-cadherin mRNA in Cag A ＋ PDCD4 plasmid group were higher than those of Cag A plasmid group（ P〈 0. 05）. The expression of Twist1 protein and Vimentin protein in Cag A plasmid group were higher than those in negative control group（ P〈 0. 05）. The expression of PDCD4 protein and E-cadherin protein in Cag A plasmid group were lower than those of negative control group（ P〈 0. 05）. The expression of Twist1 protein and Vimentin protein in Cag A ＋ PDCD4 plasmid group were lower than those in Cag A plasmid group（ P�

关 键 词：幽门螺杆菌 细胞毒素相关基因A 胃癌 程序化细胞死亡因子4 

分 类 号：R735.2[医药卫生—肿瘤]