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Immunohistochemical demonstration of EphA2 processing by MT1-MMP in invasive cutaneous squamous cell carcinoma

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Abstract

Erythropoietin-producing hepatocellular receptor-2 (EphA2) overexpression is prevalent in many types of human cancers, and it has been reported that high EphA2 expression is correlated with malignancy. Recent studies revealed that processing of EphA2 by cleaving off the N-terminal portion by membrane-type 1 matrix metalloproteinase (MT1-MMP) promotes invasion via stimulation of Ras in cancer cells in vitro. The objectives of this study were to investigate the presence and role of EphA2 processing in cutaneous squamous cell carcinoma (SCC) tissues. EphA2 (C-terminal and N-terminal) and MT1-MMP expression patterns and levels were analyzed immunohistochemically in SCC (n = 70) and Bowen disease (BD; n = 20). Levels of MT1-MMP and EphA2 expression were evaluated using digital image analysis. Proximity between MT1-MMP and EphA2 in cancer cells and its effect on EphA2 processing were investigated using a combination of in situ proximity ligation assay (PLA) and Western blotting. Immunohistochemical analyses showed that levels of EphA2 N-terminal expression were significantly lower than those of EphA2 C-terminal expression in SCC, whereas levels of EphA2 C- and N-terminal expression were similar in BD. Western blotting showed processed EphA2 fragments in human SCC tissues. Expression levels of MT1-MMP, EphA2, and processed EphA2 fragments were higher in SCC than BD. Proximity between MT1-MMP and EphA2 in SCC was demonstrated by in situ PLA. Our results suggest possible involvement of MT1-MMP processing of EphA2 in invasiveness of cutaneous SCC.

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Acknowledgments

We thank Ms. M. Onitsuka and Ms. H. Fukagawa for excellent technical assistance in immunohistochemical and in vitro studies. This work was supported in part by grants from the Research Center for Advanced Molecular Medicine, Fukuoka University and the Izumo City Supporting Cancer Research Project.

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Authors and Affiliations

  1. Department of Pathology, Fukuoka University Hospital and School of Medicine, 7-45-1 Nanakuma, Jonan-ku, Fukuoka, 814-0180, Japan

    Ryoko Tatsukawa, Kaori Koga, Mikiko Aoki & Kazuki Nabeshima

  2. Department of Dermatology, Fukuoka University Hospital, 7-45-1 Nanakuma, Jonan-ku, Fukuoka, 814-0180, Japan

    Ryoko Tatsukawa, Shinichi Imafuku & Juichiro Nakayama

  3. Research Institute, Kanagawa Cancer Center, 2-3-2 Nakao, Asahi-ku, Yokohama, 241-8515, Japan

    Naohiko Koshikawa

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  1. Ryoko Tatsukawa
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Correspondence to Kazuki Nabeshima.

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This study was approved by the Ethics Committee of Fukuoka University School of Medicine (No 14–9-09).

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Tatsukawa, R., Koga, K., Aoki, M. et al. Immunohistochemical demonstration of EphA2 processing by MT1-MMP in invasive cutaneous squamous cell carcinoma. Virchows Arch 469, 25–34 (2016). https://doi.org/10.1007/s00428-016-1934-9

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  • DOI: https://doi.org/10.1007/s00428-016-1934-9

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