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Immunohistochemical detection of bombesin receptor subtypes GRP-R and BRS-3 in human tumors using novel antipeptide antibodies

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Abstract

Bombesin (BN)-like peptides can stimulate cancer cell growth through binding to their specific G protein-coupled receptors. It is well established that BN receptors are being overexpressed in a subset of human tumors; however, little is known about the cellular and subcellular localization of individual BN receptor subtypes in these tissues. In this study, we developed and characterized novel antipeptide antibodies to the carboxy terminal regions of the gastrin-releasing peptide-preferring bombesin receptor (GRP-R) and the bombesin receptor subtype-3 (BRS-3). Specificity of the antisera was demonstrated by (1) detection of broad bands migrating at M r 50,000–70,000 in Western blots of membranes from receptor-expressing tissues; (2) cell surface staining of transfected cells; (3) translocation of GRP-R receptor immunostaining after BN exposure; and (4) abolition of tissue immunostaining by preadsorbtion of the antibodies with their immunizing peptides. The distribution of BN receptors was investigated in 74 formalin-fixed, paraffin-embedded human tumors. GRP-R receptors were most frequently detected in breast and prostate carcinomas. BRS-3 receptors were often detected in prostate and pancreatic carcinomas and in pituitary adenomas. Immunoreactive GRP-R and BRS-3 receptors were in many cases predominantly confined to the plasma membrane and uniformly present on nearly all tumor cells. The development of these novel antipeptide antibodies will facilitate the identification of those tumors, which may be targets for diagnostic or radiotherapeutic application of subtype-selective BN analogs.

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Abbreviations

BN:

bombesin

BRS-3:

bombesin receptor subtype-3

GRP:

gastrin-releasing peptide

GRP-R:

gastrin-releasing peptide receptor

NMB:

neuromedin B

NMB-R:

neuromedin B receptor

HEK:

human embryonic kidney

ir:

immunoreactivity

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Acknowledgements

We thank Beate Peter and Dana Mayer for skillful technical assistance.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Otto-von-Guericke-University, 39120, Magdeburg, Germany

    Solveig Schulz

  2. Department of Pathology, Otto-von-Guericke-University, 39120, Magdeburg, Germany

    Christoph Röcken

  3. Department of Pharmacology and Toxicology, Otto-von-Guericke-University, Leipziger Strasse 44, 39120, Magdeburg, Germany

    Stefan Schulz

  4. Department of Pathology, Charité Universitätsmedizin Berlin, Schumannstrasse 20/21, 10117, Berlin, Germany

    Christoph Röcken

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  1. Solveig Schulz
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Correspondence to Stefan Schulz.

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Schulz, S., Röcken, C. & Schulz, S. Immunohistochemical detection of bombesin receptor subtypes GRP-R and BRS-3 in human tumors using novel antipeptide antibodies. Virchows Arch 449, 421–427 (2006). https://doi.org/10.1007/s00428-006-0265-7

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  • DOI: https://doi.org/10.1007/s00428-006-0265-7

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