幽门螺杆菌毒力因子CagA和铵离子对AGS细胞中黏蛋白的影响。

Effects of the Helicobacter pylori Virulence Factor CagA and Ammonium Ion on Mucins in AGS Cells.

作者信息

Zhang Xiaoyu, Shi Ding, Liu Yong Pan, Chen Wu Jie, Wu Dong

机构信息

Department of Gastroenterology, Henan University of Chinese Medicine, Zhengzhou, China.

Department of Gastroenterology, Ningbo No. 2 Hospital, Ningbo, China.

出版信息

Yonsei Med J. 2018 Jul;59(5):633-642. doi: 10.3349/ymj.2018.59.5.633.

DOI:10.3349/ymj.2018.59.5.633

PMID:29869461

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5990679/

Abstract

PURPOSE

To investigate the effects of Helicobacter pylori (H. pylori)-CagA and the urease metabolite NH₄⁺ on mucin expression in AGS cells.

MATERIALS AND METHODS

AGS cells were transfected with CagA and/or treated with different concentrations of NH₄CL. Mucin gene and protein expression was assessed by qPCR and immunofluorescence assays, respectively.

RESULTS

CagA significantly upregulated MUC5AC, MUC2, and MUC5B expression in AGS cells, but did not affect E-cadherin and MUC6 expression. MUC5AC, MUC6, and MUC2 expression in AGS cells increased with increasing NH₄⁺ concentrations until reaching a peak level at 15 mM. MUC5B mRNA expression in AGS cells (NH₄⁺ concentration of 15 mM) was significantly higher than that at 0, 5, and 10 mM NH₄⁺. No changes in E-cadherin expression in AGS cells treated with NH₄⁺ were noted, except at 20 mM. The expression of MUC5AC, MUC2, and MUC6 mRNA in CagA-transfected AGS cells at an NH₄⁺ concentration of 15 mM was significantly higher than that at 0 mM, and decreased at higher concentrations. The expression of MUC5B mRNA increased with increases in NH₄⁺ concentration, and was significantly higher compared to that in untreated cells. No significant change in the expression of E-cadherin mRNA in CagA-transfected AGS cells was observed. Immunofluorescence assays confirmed the observed changes.

CONCLUSION

H. pylori may affect the expression of MUC5AC, MUC2, MUC5B, and MUC6 in AGS cells via CagA and/or NH₄⁺, but not E-cadherin.

摘要

目的

研究幽门螺杆菌（H. pylori）细胞毒素相关基因A（CagA）和脲酶代谢产物NH₄⁺对AGS细胞黏蛋白表达的影响。

材料与方法

用CagA转染AGS细胞和/或用不同浓度的氯化铵（NH₄CL）处理。分别通过定量聚合酶链反应（qPCR）和免疫荧光分析评估黏蛋白基因和蛋白表达。

结果

CagA显著上调AGS细胞中黏蛋白5AC（MUC5AC）、黏蛋白2（MUC2）和黏蛋白5B（MUC5B）的表达，但不影响E-钙黏蛋白和黏蛋白6（MUC6）的表达。AGS细胞中MUC5AC、MUC6和MUC2的表达随NH₄⁺浓度增加而增加，直至在15 mM时达到峰值水平。AGS细胞中MUC5B信使核糖核酸（mRNA）表达（NH₄⁺浓度为15 mM）显著高于0、5和10 mM NH₄⁺时。用NH₄⁺处理的AGS细胞中，除20 mM外，E-钙黏蛋白表达无变化。在NH₄⁺浓度为15 mM时，CagA转染的AGS细胞中MUC5AC、MUC2和MUC6 mRNA的表达显著高于0 mM时，且在更高浓度时降低。MUC5B mRNA的表达随NH₄⁺浓度增加而增加，且与未处理细胞相比显著更高。在CagA转染的AGS细胞中，未观察到E-钙黏蛋白mRNA表达的显著变化。免疫荧光分析证实了观察到的变化。