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Differential expression of tartrate-resistant acid phosphatase isoforms 5a and 5b by tumor and stromal cells in human metastatic bone disease

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Abstract

Tartrate-resistant acid phosphatase (TRAP) exists in human serum as two major isoforms, monomeric 5a and proteolytically processed enzymatically active 5b. The 5b isoform is secreted by osteoclasts and has recently been advocated as a serum marker for bone metastasis in breast cancer patients. The 5a isoform, on the other hand, is not bone-derived and has been proposed to be a marker of activated macrophages and chronic inflammation. In this study, expression of TRAP protein and enzymatic activity in bone metastases from different primary sites was examined. TRAP activity was high in bone metastases from prostate cancer, intermediate in breast cancer, and low in lung and kidney cancers. The partially purified TRAP from breast cancer bone metastasis samples exhibited the enzymatic characteristics of purple acid phosphatase. Both 5a and 5b isoforms were expressed in bone metastases of different histogenetic origins, i.e. prostate, breast, lung and kidney, and also a novel previously unreported 42 kDa variant of the TRAP 5a isoform was identified in bone metastases. This novel TRAP 5a isoform was absent in human bone, indicating that the 42 kDa variant is specific to metastatic cancer tissue. Immunohistochemistry revealed that metastatic cancer cells were the predominant source of TRAP 5a, whereas tumor-associated macrophages and occasionally multinucleated giant cells in the tumor stroma preferentially expressed the proteolytically processed TRAP 5b variant. Our results indicate the presence of a previously unstudied variant of monomeric TRAP 5a in cancer cells, which may have functional and diagnostic implications. Moreover, the presence of TRAP-positive macrophages in bone metastases could, together with cancer cells and osteoclasts, contribute to the elevated levels of serum TRAP activity observed in patients with bone metastases.

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Abbreviations

TRAP:

Tartrate-resistant acid phosphatase

OPN:

Osteopontin

BSA:

Bovine serum albumin

FPLC:

Fast protein liquid chromatography

pNPP:

p-Nitrophenylphosphate

PVDF:

Polyvinylidene fluoride

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Acknowledgments

The authors thank Dr. Jussi Halleen for kindly providing the human recombinant TRAP. This work was supported by grants from the Swedish Research Council, Cancer-Allergy Foundation, Stockholm County Council (ALF) and Karolinska Institutet Research Funds.

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Authors and Affiliations

  1. Division of Pathology F 46, Department of Laboratory Medicine, Karolinska Institute, Karolinska University Hospital, S-141 86, Huddinge, Sweden

    Serhan Zenger, Wentao He, Barbro Ek-Rylander, Daphne Vassiliou & Göran Andersson

  2. Oncology Service, Department of Orthopaedics, Karolinska University Hospital, Stockholm, Sweden

    Rickard Wedin & Henrik Bauer

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  1. Serhan Zenger
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  2. Wentao He
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  3. Barbro Ek-Rylander
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  7. Göran Andersson
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Correspondence to Serhan Zenger.

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Serhan Zenger and Wentao He have contributed equally to the study and should both be considered as first authors.

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10585_2010_9358_MOESM1_ESM.tif

Supplementary Figure 1. Anti-monomeric TRAP immunostaining on primary breast cancer tissue sections gave strong signals in cancer cells in. Magnification ×40. (TIFF 2481 kb)

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Zenger, S., He, W., Ek-Rylander, B. et al. Differential expression of tartrate-resistant acid phosphatase isoforms 5a and 5b by tumor and stromal cells in human metastatic bone disease. Clin Exp Metastasis 28, 65–73 (2011). https://doi.org/10.1007/s10585-010-9358-4

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  • DOI: https://doi.org/10.1007/s10585-010-9358-4

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