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Amino acid deletions at positions 893 and 894 of cytotoxinassociated gene A protein affect Helicobacter pylori gastric epithelial cell interactions

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World Journal of Gastroenterology 2024年第41期30卷 4449-4460页

作者： Zhi-Jing Xue Ya-Nan Gong Li-Hua He Lu Sun Yuan-Hai You Dong-Jie Fan Mao-Jun Zhang Xiao-Mei Yan Jian-Zhong Zhang Research Center of Translational Medicine Central Hospital Affiliated to Shandong First Medical UniversityJinan 250013Shandong ProvinceChina National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Communicable Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China

BACKGROUND Helicobacter pylori(***)persistently colonizes the human gastric mucosa in more than 50%of the global population,leading to various gastroduodenal diseases ranging from chronic gastritis to gastric ***-associated gene A(CagA)protein,an important oncoprotein,has highly polymorphic glu-pro-ile-tyr-ala segments at the carboxyl terminus,which play crucial roles in *** previous study revealed a significant association between amino acid deletions at positions 893 and 894 and gastric *** To investigate the impact of amino acid deletions at positions 893 and 894 on CagA *** We selected a representative HZT strain from a gastric cancer patient with amino acid deletions at positions 893 and *** cagA gene was amplified and mutated into cagA-NT and cagA-NE(sequence characteristics of strains from nongastric cancer patients),cloned and inserted into pAdtrack-CMV,and then transfected into AGS *** expression of cagA and its mutants was examined using realtime polymerase chain reaction and Western blotting,cell elongation via cell counting,F-actin cytoskeleton visualization using fluorescence staining,and interleukin-8(IL-8)secretion via enzyme-linked immunosorbent *** The results revealed that pAdtrack/cagA induced a more pronounced hummingbird phenotype than pAdtrack/cagA-NT and pAdtrack/cagA-NE(40.88±3.10 vs 32.50±3.17,P<0.001 and 40.88±3.10 vs 32.17±3.00,P<0.001)at 12 hours after *** 24 hours,pAdtrack/cagA-NE induced significantly fewer hummingbird phenotypes than pAdtrack/cagA and pAdtrack/cagA-NT(46.02±2.12 vs 53.90±2.10,P<0.001 and 46.02±2.12 vs 51.15±3.74,P<0.001).The total amount of F-actin caused by pAdtrack/cagA was significantly lower than that caused by pAdtrack/cagA-NT and pAdtrack/cagA-NE(27.54±17.37 vs 41.51±11.90,P<0.001 and 27.54±17.37 vs 41.39±14.22,P<0.001)at 12 hours after ***,pAdtrack/cagA induced higher IL-8 secretion than pAdtrack/cagA-NT a

关键词： Cytotoxin-associated gene A glu-pro-ile-tyr-ala Hummingbird phenotype Interleukin-8 Helicobacter pylori

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CagA EPIYA polymorphisms in Colombian Helicobacter pylori strains and their influence on disease-associated cellular responses

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World Journal of Gastrointestinal Oncology 2013年第3期5卷 50-59页

作者： Carlos Alberto Fajardo Andrés Javier Quiroga Andrea Coronado Karen Labrador Nicole Acosta Pilar Delgado Carlos Jaramillo María Mercedes Bravo Cancer and Infectious Agents Research Group National Cancer Institute Molecular Diagnostics and Bioinformatics Laboratory Biological Sciences Department Los Andes University

AIM: To investigate the influence of the CagA diversity in Helicobacter pylori (H. pylori ) strains from Colombia on the host cell biology. METHODS: Eighty-four H. pylori-cagA positive strains with different glu-pro-ile-tyr-ala (EPIYA) motifs patterns, isolated from patients with gastritis (n=17), atrophic gastritis (n=17), duodenal ulcer (n=16), intestinal metaplasia (n=16) and gastric cancer (n=18), were included. To determine the integrity of the cag pathogenicity island (cag PAI) we evaluated the presence of cagA, cagT, cagE, and cag10 genes by polymerase chain reaction. AGS gastric epithelial cellswere infected with each strain and assayed for translo-cation and tyrosine phosphorylation of CagA by western blot, secretion of interleukin-8 (IL-8) by enzyme-linked immuno sorbent assay after taking supernatants from cocultures and cell elongation induction. For cell elongation quantification, coculture photographs were taken and the proportion of "hummingbird" cells (>15 μm) was determined. RESULTS: Overall 72% (60/84) of the strains were found to harbor a functional cag PAI. Levels of phos-phorylated CagA were significantly higher for isolates from duodenal ulcer than the ones in strains from gas-tritis, atrophic gastritis, intestinal metaplasia and gastric cancer (49.1% ± 23.1% vs 21.1% ± 19.5%, P < 0.02; 49.1% ± 23.1% vs 26.2%±14.8%, P<0.045; 49.1% ± 23.1% vs 21.5% ± 19.5%, P<0.043 and 49.1% ± 23.1% vs 29.5% ± 27.1%, P < 0.047 respectively). We observed variable IL-8 expression levels ranging from 0 to 810 pg/mL and from 8.8 to 1442 pg/mL at 6 h and 30 h post-infection, respectively. cagPAI-defective strains did not induce detectable levels of IL-8 at 6 h post-infection. At 30 h post-infection all strains induced IL-8 expression in AGS cells, although cagPAI-defective strains induced significantly lower levels of IL-8 than strains with a functional cagPAI (57.1 ± 56.6 pg/mL vs 513.6 ± 338.6 pg/mL,P < 0.0001). We did not observe differences in the extent of cell

关键词： Helicobacter pylori cagA 3’region CagA protein Interleukin 8 Cell elongation glu-pro-ile-tyr-ala

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