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Down-regulation of miR-124/-214 in cutaneous squamous cell carcinoma mediates abnormal cell proliferation via the induction of ERK

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Abstract

Squamous cell carcinoma (SCC) is one of the most common skin cancers. Because its potential to recur and metastasize leads to a poor prognosis and significant mortality, it is necessary to develop new early diagnostic tools and new therapeutic approaches. In this study, we found protein levels of ERK1 and ERK2 were increased in SCC cell lines without changing mRNA levels and that ERK1/2 mediates abnormal cell proliferation in these cells. Then, mechanisms underlying the overexpression of ERK1/2 in SCC were investigated focusing on microRNA. We found that miR-214 is the regulator of ERK1, whereas ERK2 is regulated by miR-124 and miR-214. Expression of miR-124 and miR-214 was significantly down-regulated in SCC in vitro and in vivo. Treatment with 5-aza-deoxycytidine and trichostatin A synergistically recovered the miR-124/-214 down-regulation in SCC cell line. However, bisulphite sequencing revealed the methylation status of miR-124/-214 promoter was not increased in the SCC cell line and tumor tissue. Taken together, the down-regulation of miR-124/-214 in SCC is most likely caused, at least in part, by hypermethylation of other promoter regions rather than the miR-124/-214 promoter. Supplementation of these microRNAs in the SCC cell line reduced the abnormal cell proliferation by normalizing ERK1/2 levels. Additionally, serum concentration of miR-124 was correlated with miR-124 expression levels in the tumor tissues and inversely correlated with tumor progression. On the other hand, miR-214 was not detected in the serum. Investigation of the regulatory mechanisms of keratinocyte proliferation by microRNA may lead to develop new biomarkers and treatments using microRNA.

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Acknowledgments

We thank Ms. Junko Suzuki and Ms. Chiemi Shiotsu for their valuable technical assistance. We also would like to thank Dr. Norbert E. Fusenig for kindly providing HaCaT.

Disclosure statement

The authors declare that they have no competing financial interests that might influence the results and discussion in this paper.

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Authors and Affiliations

  1. Department of Dermatology and Plastic Surgery, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto, Japan

    Keitaro Yamane, Masatoshi Jinnin, Tomomi Etoh, Yuki Kobayashi, Naoki Shimozono, Satoshi Fukushima, Shinichi Masuguchi, Keishi Maruo, Yuji Inoue, Tsuyoshi Ishihara, Jun Aoi & Hironobu Ihn

  2. Department of Molecular Genetics, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto, Japan

    Jun Aoi & Yuichi Oike

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  1. Keitaro Yamane
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  2. Masatoshi Jinnin
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Correspondence to Masatoshi Jinnin.

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Yamane, K., Jinnin, M., Etoh, T. et al. Down-regulation of miR-124/-214 in cutaneous squamous cell carcinoma mediates abnormal cell proliferation via the induction of ERK. J Mol Med 91, 69–81 (2013). https://doi.org/10.1007/s00109-012-0935-7

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  • DOI: https://doi.org/10.1007/s00109-012-0935-7

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