BARX2 expression is downregulated by CpG island hypermethylation and is associated with suppressed cell proliferation and invasion of gastric cancer cells

Ma,Juan; Xia,Ling‑Ling; Yao,Xue‑Qing; Zheng,Shi‑Min; Li,Shi; Xu,Li‑Shu; Sha,Wei‑Hong; Li,Ze‑Song

doi:10.3892/or.2020.7558

BARX2 expression is downregulated by CpG island hypermethylation and is associated with suppressed cell proliferation and invasion of gastric cancer cells

Authors:
- Juan Ma
- Ling‑Ling Xia
- Xue‑Qing Yao
- Shi‑Min Zheng
- Shi Li
- Li‑Shu Xu
- Wei‑Hong Sha
- Ze‑Song Li
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Affiliations: Department of Gastroenterology and Hepatology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, and Guangdong Provincial Geriatrics Institute, Guangzhou, Guangdong 510080, P.R. China, Guangdong Key Laboratory of Systems Biology and Synthetic Biology for Urogenital Tumors, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, Guangdong 518000, P.R. China, Department of General Surgery, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510080, P.R. China
Published online on: March 22, 2020 https://doi.org/10.3892/or.2020.7558
Pages: 1805-1818
Copyright: © Ma et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

BarH‑like homeobox 2 (BARX2), a homeobox gene, is associated with several types of cancers. The present study aimed to determine whether DNA methylation downregulates BARX2 expression and whether BARX2 is associated with suppression of gastric carcinogenesis. BARX2 protein expression in normal and cancerous gastric tissues and various gastric cancer (GC) cell lines was detected using immunohistochemical and western blot assays. BARX2 mRNA levels were detected using both reverse transcription‑polymerase chain reaction (RT‑PCR) and quantitative PCR (qPCR). Promoter hypermethylation in GC cells was detected using methylation‑specific PCR or bisulfite DNA sequencing PCR. Effects of BARX2 expression on GC cell proliferation, clonal formation, and migration were evaluated after lentivirus‑BARX2 transfection. The effect of stable BARX2 transfection on tumor formation was assessed in a nude xenograft mouse model. BARX2 was strongly expressed in the normal gastric mucosa, but weakly or not expressed in GC tissues and most GC cell lines. BARX2 expression was negatively correlated with DNMT (a marker for DNA methylation) expression in the gastric tissues. The BARX2 promoter fragment was hypermethylated in the GC cell lines. Overexpression of BARX2 significantly inhibited GC cell proliferation, clonal formation, and migration. Stable BARX2 transfection inhibited tumor formation in xenograft mice, which was correlated with decreased expression of E‑cadherin, proliferation markers, and matrix metalloproteinases. In conclusion, BARX2 expression is aberrantly reduced in GC, which is associated with increased DNA methylation of its promoter. BARX2 inhibits GC cell proliferation, migration, and tumor formation, suggesting that BARX2 acts as a tumor suppressor in gastric carcinogenesis.

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