Effects of Helicobacter pylori on the expression levels of GATA‑3 and connexin 32 and the GJIC function in gastric epithelial cells and their association by promoter analysis

Huang,Lihua; Guo,Yinjie; Cao,Dan; Liu,Xiaoming; Zhang,Linfang; Cao,Ke; Hu,Tingzi; Qi,Yong; Xu,Canxia

doi:10.3892/ol.2018.8796

Effects of Helicobacter pylori on the expression levels of GATA‑3 and connexin 32 and the GJIC function in gastric epithelial cells and their association by promoter analysis

Authors:
- Lihua Huang
- Yinjie Guo
- Dan Cao
- Xiaoming Liu
- Linfang Zhang
- Ke Cao
- Tingzi Hu
- Yong Qi
- Canxia Xu
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Affiliations: Center for Medical Experiments, Third Xiangya Hospital of Central South University, Changsha, Hunan 410013, P.R. China, Department of Gastroenterology, Third Xiangya Hospital of Central South University, Changsha, Hunan 410013, P.R. China, Department of Oncology, Third Xiangya Hospital of Central South University, Changsha, Hunan 410013, P.R. China, Clinical Laboratory, Third Xiangya Hospital of Central South University, Changsha, Hunan 410013, P.R. China
Published online on: May 24, 2018 https://doi.org/10.3892/ol.2018.8796
Pages: 1650-1658
Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to explore the effects of Helicobacter pylori (H. pylori) infection on the expression of transcription factor GATA binding protein 3 (GATA‑3) and connexin 32 (Cx32) in cultured gastric mucosa cells, and their association with each other. GES‑1 cells were co‑cultured with East Asian type cytotoxin‑associated gene A+ H. pylori in the H. pylori group, and without H. pylori culture in the control group. Additionally, Mongolian gerbils were gavaged with H. pylori, and later the gastric antrum tissues were collected. The GATA‑3 and Cx32 mRNA and protein expression levels were detected by a reverse transcription‑quantitative polymerase chain reaction and western blot analysis, respectively. The scratch labeling fluorescent dye tracer (SLDT) technique was used to detect the gap junctional intercellular communication (GJIC) function. GATA‑3 small interfering RNA (siRNA) was transfected into BGC823 cells and its effect on Cx32 expression levels was detected. The impact of GATA‑3 on Cx32 promoter transcriptional activity was detected using a dual luciferase reporter assay. The results revealed that H. pylori infection increased GATA‑3 expression and decreased Cx32 expression in GES‑1 cells and in animal gastric tissues compared with their respective controls, whilst in BGC823 cells, GATA‑3 siRNA increased Cx32 expression compared with the control. In the SLDT experiment of GES‑1 cells with H. pylori infection, the fluorescent dye was primarily limited to a single cell row close to the scratch, and only a limited amount of dye passing to the second cell row, indicating that the GJIC function was substantially reduced or absent compared with the control group, where the fluorescence dye transferred to the neighboring cells of 3‑4 rows, indicating a stronger GJIC function comparatively. GATA‑3 inhibited the expression of the luciferase reporter gene, compared with the controls, suggesting that GATA‑3 inhibited the expression of Cx32 by binding to Cx32 promoter sites. These results indicated that H. pylori‑increased GATA‑3 expression, which downregulated Cx32 expression, may serve an important function in gastric carcinogenesis, and GATA‑3 siRNA may serve a function in the prevention and treatment of gastric cancer.

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