Helicobacter pylori infection impairs gastric epithelial barrier function via microRNA‑100‑mediated mTOR signaling inhibition

Hu,Guimei; Guo,Lihua; Ye,Guoliang

doi:10.3892/mmr.2018.8971

Helicobacter pylori infection impairs gastric epithelial barrier function via microRNA‑100‑mediated mTOR signaling inhibition

Authors:
- Guimei Hu
- Lihua Guo
- Guoliang Ye
View Affiliations

Affiliations: Department of Gastroenterology, The Affiliated Hospital of Medical School of Ningbo University, Ningbo, Zhejiang 315020, P.R. China
Published online on: May 4, 2018 https://doi.org/10.3892/mmr.2018.8971
Pages: 587-594

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Helicobacter pylori (H. pylori) infection has an important effect on human health as it is an established cause of gastric carcinoma. microRNAs (miRNAs/miRs) are a family of small RNAs with various functions in the control of cellular profiles. However, the effect of miR‑100 in H. pylori infection remains unknown. Healthy volunteers (n=100) and patients with H. pylori infection (n=98) were included in the present study. H. pylori infection was confirmed by urea breath tests. The levels of miR‑100 in gastroscopic biopsy samples and cultured GES‑1 cells were measured by reverse transcription‑quantitative polymerase chain reaction. Furthermore, miR‑100 was overexpressed or inhibited in GES‑1 cells by an miR‑100 mimic or inhibitor, respectively. The expression of cell‑junction proteins and members of the mechanistic target of rapamycin kinase (mTOR) signaling pathway was investigated by western blotting. The results demonstrated that miR‑100 levels were upregulated in infected patients and cultured gastric epithelial cells, compared with the respective controls. Additionally, the expression of epithelial (E)‑cadherin and zona occludens‑1 in the gastric mucosa of infected patients and GES‑1 cells was downregulated. Furthermore, infected gastric epithelial cells exhibited impaired barrier functions, as measured by resistance and permeability tests. Overexpression of miR‑100 inhibited junction protein expression, as well as the activation of the mTOR signaling pathway, while suppression of miR‑100 restored E‑cadherin expression and mTOR signaling. The results of the present study indicate that H. pylori infection may cause dysfunction of the gastric epithelial barrier by increasing miR‑100 levels, which subsequently inhibit mTOR signaling. These results may have potential applications affecting miR‑100 in H. pylori‑related diseases.

View Figures

View References

1	Warren JR and Marshall B: Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet. 1:1273–1275. 1983.PubMed/NCBI
2	Everhart JE: Recent developments in the epidemiology of Helicobacter pylori. Gastroenterol Clin North Am. 29:559–578. 2000. View Article : Google Scholar : PubMed/NCBI
3	McColl KE: Clinical practice. Helicobacter pylori infection. N Engl J Med. 362:1597–1604. 2010. View Article : Google Scholar : PubMed/NCBI
4	Censini S, Lange C, Xiang Z, Crabtree JE, Ghiara P, Borodovsky M, Rappuoli R and Covacci A: cag, a pathogenicity island of Helicobacter pylori, encodes type I-specific and disease-associated virulence factors. Proc Natl Acad Sci USA. 93:14648–14653. 1996. View Article : Google Scholar : PubMed/NCBI
5	Selbach M, Moese S, Hauck CR, Meyer TF and Backert S: Src is the kinase of the Helicobacter pylori CagA protein in vitro and in vivo. J Biol Chem. 277:6775–6778. 2002. View Article : Google Scholar : PubMed/NCBI
6	Tammer I, Brandt S, Hartig R, König W and Backert S: Activation of Abl by Helicobacter pylori: A novel kinase for CagA and crucial mediator of host cell scattering. Gastroenterology. 132:1309–1319. 2007. View Article : Google Scholar : PubMed/NCBI
7	Backert S, Moese S, Selbach M, Brinkmann V and Meyer TF: Phosphorylation of tyrosine 972 of the Helicobacter pylori CagA protein is essential for induction of a scattering phenotype in gastric epithelial cells. Mol Microbiol. 42:631–644. 2001. View Article : Google Scholar : PubMed/NCBI
8	Mueller D, Tegtmeyer N, Brandt S, Yamaoka Y, De Poire E, Sgouras D, Wessler S, Torres J, Smolka A and Backert S: c-Src and c-Abl kinases control hierarchic phosphorylation and function of the CagA effector protein in Western and East Asian Helicobacter pylori strains. J Clin Invest. 122:1553–1566. 2012. View Article : Google Scholar : PubMed/NCBI
9	Saadat I, Higashi H, Obuse C, Umeda M, Murata-Kamiya N, Saito Y, Lu H, Ohnishi N, Azuma T, Suzuki A, et al: Helicobacter pylori CagA targets PAR1/MARK kinase to disrupt epithelial cell polarity. Nature. 447:330–333. 2007. View Article : Google Scholar : PubMed/NCBI
10	Palframan SL, Kwok T and Gabriel K: Vacuolating cytotoxin A (VacA), a key toxin for Helicobacter pylori pathogenesis. Front Cell Infect Microbiol. 2:922012. View Article : Google Scholar : PubMed/NCBI
11	Jain P, Luo ZQ and Blanke SR: Helicobacter pylori vacuolating cytotoxin A (VacA) engages the mitochondrial fission machinery to induce host cell death. Proc Natl Acad Sci USA. 108:16032–16037. 2011. View Article : Google Scholar : PubMed/NCBI
12	Rassow J and Meinecke M: Helicobacter pylori VacA: A new perspective on an invasive chloride channel. Microbes Infect. 14:1026–1033. 2012. View Article : Google Scholar : PubMed/NCBI
13	Boquet P and Ricci V: Intoxication strategy of Helicobacter pylori VacA toxin. Trends Microbiol. 20:165–174. 2012. View Article : Google Scholar : PubMed/NCBI
14	Lagos-Quintana M, Rauhut R, Lendeckel W and Tuschl T: Identification of novel genes coding for small expressed RNAs. Science. 294:853–858. 2001. View Article : Google Scholar : PubMed/NCBI
15	Bartel DP: MicroRNAs: Target recognition and regulatory functions. Cell. 136:215–233. 2009. View Article : Google Scholar : PubMed/NCBI
16	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
17	Nagaraja AK, Creighton CJ, Yu Z, Zhu H, Gunaratne PH, Reid JG, Olokpa E, Itamochi H, Ueno NT, Hawkins SM, et al: A link between mir-100 and FRAP1/mTOR in clear cell ovarian cancer. Mol Endocrinol. 24:447–463. 2010. View Article : Google Scholar : PubMed/NCBI
18	Li BH, Zhou JS, Ye F, Cheng XD, Zhou CY, Lu WG and Xie X: Reduced miR-100 expression in cervical cancer and precursors and its carcinogenic effect through targeting PLK1 protein. Eur J Cancer. 47:2166–2174. 2011. View Article : Google Scholar : PubMed/NCBI
19	Leite KR, Sousa-Canavez JM, Reis ST, Tomiyama AH, Camara-Lopes LH, Sañudo A, Antunes AA and Srougi M: Change in expression of miR-let7c, miR-100 and miR-218 from high grade localized prostate cancer to metastasis. Urol Oncol. 29:265–269. 2011. View Article : Google Scholar : PubMed/NCBI
20	Wei Y, Wang T, Song H, Tian L, Lyu G, Zhao L and Xue Y: C-C motif chemokine 22 ligand (CCL22) concentrations in sera of gastric cancer patients are related to peritoneal metastasis and predict recurrence within one year after radical gastrectomy. J Surg Res. 211:266–278. 2017. View Article : Google Scholar : PubMed/NCBI
21	Temmesfeld-Wollbrück B, Brell B, zu Dohna C, Dorenberg M, Hocke AC, Martens H, Klar J, Suttorp N and Hippenstiel S: Adrenomedullin reduces intestinal epithelial permeability in vivo and in vitro. Am J Physiol Gastrointest Liver Physiol. 297:G43–G51. 2009. View Article : Google Scholar : PubMed/NCBI
22	Oshima T, Miwa H and Joh T: Aspirin induces gastric epithelial barrier dysfunction by activating p38 MAPK via claudin-7. Am J Physiol Cell Physiol. 295:C800–C806. 2008. View Article : Google Scholar : PubMed/NCBI
23	Franco AT, Israel DA, Washington MK, Krishna U, Fox JG, Rogers AB, Neish AS, Collier-Hyams L, Perez-Perez GI, Hatakeyama M, et al: Activation of beta-catenin by carcinogenic Helicobacter pylori. Proc Natl Acad Sci USA. 102:10646–10651. 2005. View Article : Google Scholar : PubMed/NCBI
24	Hou J, Wang P, Lin L, Liu X, Ma F, An H, Wang Z and Cao X: MicroRNA-146a feedback inhibits RIG-I-dependent Type I IFN production in macrophages by targeting TRAF6, IRAK1, and IRAK2. J Immunol. 183:2150–2158. 2009. View Article : Google Scholar : PubMed/NCBI
25	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
26	Hartsock A and Nelson WJ: Adherens and tight junctions: Structure, function and connections to the actin cytoskeleton. Biochim Biophys Acta. 1778:660–669. 2008. View Article : Google Scholar : PubMed/NCBI
27	Sampson LL, Davis AK, Grogg MW and Zheng Y: mTOR disruption causes intestinal epithelial cell defects and intestinal atrophy postinjury in mice. FASEB J. 30:1263–1275. 2016. View Article : Google Scholar : PubMed/NCBI
28	Kaser A and Blumberg RS: Autophagy, microbial sensing, endoplasmic reticulum stress, and epithelial function in inflammatory bowel disease. Gastroenterology. 140:1738–1747. 2011. View Article : Google Scholar : PubMed/NCBI
29	Ajani JA, Lee J, Sano T, Janjigian YY, Fan D and Song S: Gastric adenocarcinoma. Nat Rev Dis Primers. 3:170362017. View Article : Google Scholar : PubMed/NCBI
30	de Martel C, Ferlay J, Franceschi S, Vignat J, Bray F, Forman D and Plummer M: Global burden of cancers attributable to infections in 2008: A review and synthetic analysis. Lancet Oncol. 13:607–615. 2012. View Article : Google Scholar : PubMed/NCBI
31	Polk DB and Peek RM Jr: Helicobacter pylori: Gastric cancer and beyond. Nat Rev Cancer. 10:403–414. 2010. View Article : Google Scholar : PubMed/NCBI
32	Grivennikov SI, Greten FR and Karin M: Immunity, inflammation, and cancer. Cell. 140:883–899. 2010. View Article : Google Scholar : PubMed/NCBI
33	Croce CM: Causes and consequences of microRNA dysregulation in cancer. Nat Rev Genet. 10:704–714. 2009. View Article : Google Scholar : PubMed/NCBI