Cag A阳性幽门螺杆菌对胃癌细胞lncRNA DLEU2表达及上皮间质转化的影响  

Effects of Cag A positive Hp on lncRNA DLEU2 expression and epithelial mesenchymal transformation of gastric cancer cells

作　　者：曲雪 刘琦 赵鑫 邢元欣 郏雁飞 郑燕 杨炜华 汪运山 韩淑毅 QU Xue;LIU Qi;ZHAO Xin;XING Yuanxin;JIA Yanfei;ZHENG Yan;YANG Weihua;WANG Yunshan;HAN Shuyi(Graduate School,Shandong First Medical University(Shandong Academy of Medical Sciences),Jinan 250117,China;不详)

机构地区：[1]山东第一医科大学(山东省医学科学院)研究生院,济南250117 [2]山东大学齐鲁医学院 [3]山东第一医科大学附属中心医院医学实验诊断中心 [4]山东第一医科大学附属中心医院基础医学研究中心 [5]山东第一医科大学附属中心医院专科转化研究中心

出　　处：《山东医药》2023年第1期1-5,共5页Shandong Medical Journal

基　　金：山东省自然科学基金资助项目(ZR2020MH320)。

摘　　要：目的探讨细胞毒素相关蛋白A(Cag A)阳性幽门螺杆菌(Hp)对胃癌细胞长链非编码RNA淋巴细胞白血病缺失基因2(DLEU2)表达及上皮间质转化的影响。方法体外培养胃癌细胞系HGC-27、AGS、MGC-803、MKN-28以及正常胃黏膜上皮细胞系GES1,采用RT-qPCR法检测DLEU2表达,并选择DLEU2表达最高的胃癌细胞进行后续实验。将Hp NCTC标准菌株26695或11637分别与正常胃黏膜上皮细胞和胃癌细胞共培养0、3、9、12 h,采用RT-qPCR法检测DLEU2表达。采用Western blotting法检测与NCTC 26695或11637共培养3、9、12 h正常胃黏膜上皮细胞和胃癌细胞Cag A表达以及共培养3、9、12 h神经钙黏着蛋白(N-cadherin)、波形蛋白(Vimentin)表达。同时,分析与NCTC 26695或NCTC 11637共培养的胃癌细胞DLEU2、Cag A表达与N-cadherin、Vimentin表达的关系。结果HGC-27、AGS、MGC-803、MKN-28细胞DLEU2相对表达量均高于GES1细胞(P均<0.05)。在胃癌细胞中,以HGC-27细胞DLEU2相对表达量最高。因此,选择胃癌细胞HGC-27进行后续实验。NCTC 11637感染3、9、12 h时GES1细胞DLEU2相对表达量均高于NCTC 11637感染0 h时(P均<0.05),NCTC 11637感染12 h时HGC-27细胞DLEU2相对表达量高于NCTC 11637感染0 h时(P<0.05)。NCTC 11637感染12 h时HGC-27细胞和GES1细胞Cag A、N-cadherin、Vimentin相对表达量均高于NCTC 26695感染12 h时相应细胞(P均<0.05)。Pearson相关分析显示,NCTC 26695或11637感染的HGC-27细胞DLEU2、Cag A表达与N-cadherin、Vimentin表达均呈正相关关系(P均<0.05)。结论Cag A阳性Hp能够引起胃癌细胞长链非编码RNA DLEU2异常表达,而DLEU2异常表达可促进胃癌细胞上皮间质转化。Objective To investigate the effects of cytotoxin-associated protein A(Cag A)positive Helicobacter pylori(Hp)on the expression of long non-coding RNA lymphocytic leukemia deletion gene 2(DLEU2)and epithelial mesenchymal transformation of gastric cancer cells.Methods Human gastric cancer cell lines HGC-27,AGS,MGC-803,MKN-28 and human normal gastric mucosal epithelial cell line GES1 were cultured in vitro.DLEU2 expression was detected by RT-qPCR,and gastric cancer cells with the highest DLEU2 expression were selected for subsequent experiments.Hp National Collection of Type Cultures(NCTC)standard strains 26695 or 11637 were co-cultured with normal gastric epithelial cells and gastric cancer cells for 0,3,9,and 12 h,respectively.DLEU2 expression was detected by RT-qPCR.Western blotting was used to detect the expression of Cag A protein in human normal gastric epithelial cells and gastric cancer cells cultured with NCTC 26695 or 11637 for 3,9 and 12 h,and the expression of N-cadherin and Vimentin at 3,9 and 12 h.Meanwhile,we analyzed the relationships between DLEU2,Cag A expression and N-cadherin,Vimentin expression in the gastric cancer cells co-cultured with NCTC 26695 or 11637 strains.Results The relative expression of DLEU2 in the HGC-27,AGS,MGC-803,MKN-28 cells was higher than that in the GES1 cells(P<0.05).In the human gastric cancer cells,HGC-27 cells had the highest relative expression of DLEU2.Therefore,human gastric cancer cells HGC-27were selected for subsequent experiments.The relative expression of DLEU2 in the GES1 cells infected with NCTC 11637for 3,9 and 12 h was higher than that in GES1 cells infected with sterile strains for 0 h(P<0.05).The relative expression of DLEU2 in the HGC-27 cells infected with NCTC 11637 for 12 h was higher than that in the HGC-27 cells infected with sterile strains for 0 h(P<0.05).The relative expression of Cag A protein in the HGC-27 and GES1 cells infected with NCTC 11637 strain at 3,9,12 h was higher than that in the HGC-27 and GES1 cells infected with NCTC 26695 strain

关 键 词：胃癌 幽门螺杆菌 细胞毒素相关蛋白A 长链非编码RNA 淋巴细胞白血病缺失基因2 上皮间质转化 

分 类 号：R735.2[医药卫生—肿瘤]