Abstract
Chronic inflammation contributes to the development of various forms of cancer. The polyamine catabolic enzyme spermine oxidase (SMOX) is induced in chronic inflammatory conditions, including Helicobacter pylori-associated gastritis, where its production of hydrogen peroxide contributes to DNA damage and subsequent tumorigenesis. MicroRNA expression levels are also altered in inflammatory conditions; specifically, the tumor suppressor miR-124 becomes silenced by DNA methylation. We sought to determine if this repression of miR-124 is associated with elevated SMOX activity and concluded that miR-124 is indeed a negative regulator of SMOX. In gastric adenocarcinoma cells harboring highly methylated and silenced mir-124 gene loci, 5-azacytidine treatment allowed miR-124 re-expression and decreased SMOX expression. Overexpression of an exogenous miR-124-3p mimic repressed SMOX mRNA and protein expression as well as H2O2 production by >50% within 24 h. Reporter assays indicated that direct interaction of miR-124 with the 3′-untranslated region of SMOX mRNA contributes to this negative regulation. Importantly, overexpression of miR-124 before infection with H. pylori prevented the induction of SMOX believed to contribute to inflammation-associated tumorigenesis. Compelling human in vivo data from H. pylori-positive gastritis tissues indicated that the mir-124 gene loci are more heavily methylated in a Colombian population characterized by elevated SMOX expression and a high risk for gastric cancer. Furthermore, the degree of mir-124 methylation significantly correlated with SMOX expression throughout the population. These results indicate a protective role for miR-124 through the inhibition of SMOX-mediated DNA damage in the etiology of H. pylori-associated gastric cancer.
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Acknowledgements
We thank Alberto Delgado for his technical expertise and performance of SMOX immunohistochemistry studies. This study was supported by National Institutes of Health Grants R01CA051085 and R01CA098454 (to RAC), R01DK053620 and R01CA190612 (to KTW), P01CA028842 (to PC and KTW), P01CA116087 (to KTW), K01AT007324 (to RC), the Samuel Waxman Cancer Research Foundation (to RAC), the Vanderbilt Digestive Disease Research Center grant (P30DK058404), UL1RR024975 (Vanderbilt CTSA, Pilot Project to KTW) and Merit Review Grant 1I01BX001453 from the Office of Medical Research, Department of Veterans Affairs (to KTW).
Author contributions
TMS, KTW and RAC designed the research; TMS, JCS, MBP, RC and BGS performed the research; LEB and PC provided unique human samples; TMS, MBP, RMM, KTW and RAC analyzed the data; and TMS, KTW and RAC wrote the paper. All authors revised and approved the final manuscript.
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Murray-Stewart, T., Sierra, J., Piazuelo, M. et al. Epigenetic silencing of miR-124 prevents spermine oxidase regulation: implications for Helicobacter pylori-induced gastric cancer. Oncogene 35, 5480–5488 (2016). https://doi.org/10.1038/onc.2016.91
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DOI: https://doi.org/10.1038/onc.2016.91
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