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Development of Novel Monoclonal Antibodies for Evaluation of Transmembrane Prostate Androgen-Induced Protein 1 (TMEPAI) Expression Patterns in Gastric Cancer

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Pathology & Oncology Research

Abstract

Transmembrane prostate androgen-induced protein 1 (TMEPAI) is a single-span membrane protein, functionally involved in transforming growth factor beta signaling pathway. The particular protein presented in cells in three isoforms, which differs in the length of the soluble N-terminal extracellular domain, making it challenging for the immunochemical recognition. By using quantitative real-time polymerase chain reaction, we identified significant upregulation of PMEPA1 gene expression in malignant tissues of patients with gastric adenocarcinoma. The main part of commercially available anti-TMEPAI antibodies are having polyclonal nature or not suitable for immunocytochemical localization of target protein in tissue specimens. Hence, we decide to generate a set of novel rat monoclonal antibodies (mAb) directed against conservative C-terminal cytoplasmic epitope. Immunoblotting analysis showed that monoclonal antibodies, 2E1, 6C6, and 10A7 were able to recognize specifically target protein in transiently transfected HEK293T and CHO-K1 cells. Especially established mAb, named 10A7, showed the excellent binding ability to target protein in immunohistochemistry. By using developed antibodies, we observed pronounced expression of TMEPAI in normal gastric epithelial cells while tumor cells from gastric adenomas, and adenocarcinoma samples were mostly negative for target protein expression. Also, we found that gastric epithelium cells lose the TMEPAI expression concurrently with severe dysplasia progression, which probably caused by a mechanism involving specific microRNA.

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Acknowledgements

Authors gratefully acknowledge Prof. M Kato (University of Tsukuba, Japan) for providing original plasmid DNA. Also, an excellent technical assistance from Dr. I. V. Stepanov and Dr. M. A. Buldakov (Tomsk Cancer Research Institute) is greatly appreciated.

Author information

Author notes

  1. Mikhail S. Karbyshev, Evgeniya S. Grigorieva and Viktor V. Volkomorov contributed equally to this work

Authors and Affiliations

  1. Department of Molecular Oncology and Immunology, Cancer Research Institute, Tomsk National Research Medical Center, Russian Academy of Sciences, Kooperativny Str. 5, Tomsk, Russian Federation, 634050

    Mikhail S. Karbyshev, Evgeniya S. Grigoryeva, Viktor V. Volkomorov, Irina V. Mitrofanova, Nadezda V. Cherdyntseva & Evgeny L. Choynzonov

  2. Division of Research and Development Management, JSC «National Immunobiological Company», Kapranova Street 3, Moscow, Russian Federation, 123242

    Mikhail S. Karbyshev

  3. Department of Biochemistry and Molecular Biology, The Pirogov Russian National Research Medical University, Ostrovitianov str. 1, Moscow, Russian Federation, 117997

    Mikhail S. Karbyshev

  4. Helmholtz Zentrum München, German Research Center for Environmental Health, Institute of Molecular Immunology, Marchioninistr. 25, 81377, Munich, Germany

    Elisabeth Kremmer & Alexander Huber

  5. Laboratory for Translational Cellular and Molecular Biomedicine, Tomsk State University, 36 Lenin Ave, Tomsk, Russian Federation, 634050

    Irina V. Mitrofanova, Marina V. Zavyalova, Julia G. Kzhyshkowska & Nadezda V. Cherdyntseva

  6. Department of Pathological Anatomy and Cytology, Cancer Research Institute, Tomsk National Research Medical Center, Russian Academy of Sciences, Kooperativny Str. 5, Tomsk, Russian Federation, 634050

    Evgeniya V. Kaigorodova & Marina V. Zavyalova

  7. Department of Pathological Anatomy, Siberian State Medical University, 2 Moskovsky trakt, Tomsk, Russian Federation, 634050

    Marina V. Zavyalova

  8. Institute of Transfusion Medicine and Immunology, Medical Faculty Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany

    Julia G. Kzhyshkowska

  9. Red Cross Blood Service Baden-Württemberg–Hessen, Friedrich-Ebert Str. 107, D-68167, Mannheim, Germany

    Julia G. Kzhyshkowska

Authors
  1. Mikhail S. Karbyshev
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  2. Evgeniya S. Grigoryeva
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  3. Viktor V. Volkomorov
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  4. Elisabeth Kremmer
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  5. Alexander Huber
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  6. Irina V. Mitrofanova
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  7. Evgeniya V. Kaigorodova
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  8. Marina V. Zavyalova
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  9. Julia G. Kzhyshkowska
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  10. Nadezda V. Cherdyntseva
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  11. Evgeny L. Choynzonov
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Corresponding author

Correspondence to Evgeniya S. Grigoryeva.

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The authors declare that they have no conflict of interest.

Funding

The presented study financially supported by grants from the Russian Fund for Basic Research (14–04-31500) and Tomsk State University Competitiveness Improvement Program.

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Karbyshev, M.S., Grigoryeva, E.S., Volkomorov, V.V. et al. Development of Novel Monoclonal Antibodies for Evaluation of Transmembrane Prostate Androgen-Induced Protein 1 (TMEPAI) Expression Patterns in Gastric Cancer. Pathol. Oncol. Res. 24, 427–438 (2018). https://doi.org/10.1007/s12253-017-0247-x

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