miR-200c regulates the proliferation, apoptosis and invasion of gastric carcinoma cells through the downregulation of EDNRA expression

Wei,Wei; Shi,Li; Chen,Weichang; Hu,Liyu; Chen,Dan; Shi,Xiaoying; Xiang,Hua; Guo,Cunguo; Wu,Zhenping

doi:10.3892/ijmm.2017.3317

miR-200c regulates the proliferation, apoptosis and invasion of gastric carcinoma cells through the downregulation of EDNRA expression

Authors:
- Wei Wei
- Li Shi
- Weichang Chen
- Liyu Hu
- Dan Chen
- Xiaoying Shi
- Hua Xiang
- Cunguo Guo
- Zhenping Wu
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Affiliations: Department of Gastroenterology, Jinhua Central Hospital, Jinhua, Zhejiang 321000, P.R. China, Department of Gastroenterology, Jinhua People's Hospital, Jinhua, Zhejiang 321000, P.R. China, Department of Gastroenterology, The First Affiliated Hospital of Suzhou University, Suzhou, Jiangsu 215006, P.R. China
Published online on: December 12, 2017 https://doi.org/10.3892/ijmm.2017.3317
Pages: 1619-1626

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Abstract

This study aimed to investigate the regulatory effects of microRNA (miR)‑200c on the proliferation, apoptosis and invasion of gastric carcinoma cells and to elucidate the underlying mechanisms involving the possible role of endothelin receptor A (EDNRA). The expression levels of miR‑200c and EDNRA in the gastric carcinoma cell lines, BGC‑823, SGC‑7901 and HGC‑27, and in GES‑1 normal gastric cells were evaluated by RT‑PCR and western blot analysis. The gastric carcinoma cells, particularly the BGC‑823 cells, expressed significantly lower levels of miR‑200c than the normal gastric cells (P<0.01). Thus, the BGC‑823 cells were employed as model cells. In comparison to the normal gastric cells, EDNRA was overexpressed in the gastric carcinoma cells (P<0.01). Following the transfection of the gastric carcinoma cells with miR‑200c mimics, or negative control vector (miR‑200c NC), or with siRNA targeting EDNRA (siRNA EDNRA) or negatvie control siRNA (siRNA NC), the expression levels were assessed again by RT‑PCR and western blot analysis. The successful transfection of miR‑200c mimics was observed and this markedly elevated the expression of miR‑200c (P<0.01). The transfection of miR‑200c mimics or siRNA EDNRA notably decreased the EDNRA mRNA and protein expression levels (both P<0.01). In additoin, dual‑luciferase reporter gene analysis was performed to reveal the targeting relationship between miR‑200c and EDNRA. EDNRA was found to be the downstream target gene of miR‑200c. Moreover, methyl thiazolyl tetrazolium (MTT) assay, Hoechst staining and Transwell assay were conducted to demonstrate the effects of miR‑200c mimics or siRNA EDNRA on the proliferation, apoptosis and invasion of the gastric carcinoma cells, respectively. We found that transfection with miR‑200c mimics and siRNA EDNRA were able to markedly suppress the proliferation and invasive capacity, and to promote the apoptosis of the gastric carcinoma cells (all P<0.01). On the whole, our data indicate that miR‑200c regulates the proliferation, apoptosis and invasion of gastric carcinoma cells through the downregulation of EDNRA expression.

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