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Clinical & Experimental Metastasis

, Volume 28, Issue 1, pp 65–73 | Cite as

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

  • Serhan Zenger
  • Wentao He
  • Barbro Ek-Rylander
  • Daphne Vassiliou
  • Rickard Wedin
  • Henrik Bauer
  • Göran Andersson
Research Paper

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.

Keywords

Tartrate-resistant acid phosphatase Metastatic bone disease Macrophage Cancer Bone resorption Osteoclast 

Abbreviations

TRAP

Tartrate-resistant acid phosphatase

OPN

Osteopontin

BSA

Bovine serum albumin

FPLC

Fast protein liquid chromatography

pNPP

p-Nitrophenylphosphate

PVDF

Polyvinylidene fluoride

Notes

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.

Supplementary material

10585_2010_9358_MOESM1_ESM.tif (2.4 mb)
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)

References

  1. 1.
    Clark PE, Torti FM (2003) Prostate cancer and bone metastases: medical treatment. Clin Orthop Relat Res: S148-157Google Scholar
  2. 2.
    Roodman GD (2004) Mechanisms of bone metastasis. N Engl J Med 350:1655–1664CrossRefPubMedGoogle Scholar
  3. 3.
    El-Tanani MK (2008) Role of osteopontin in cellular signaling and metastatic phenotype. Front Biosci 13:4276–4284CrossRefPubMedGoogle Scholar
  4. 4.
    Hayman AR, Bune AJ, Bradley JR, Rashbass J, Cox TM (2000) Osteoclastic tartrate-resistant acid phosphatase (Acp 5): its localization to dendritic cells and diverse murine tissues. J Histochem Cytochem 48:219–228PubMedGoogle Scholar
  5. 5.
    Hayman AR, Macary P, Lehner PJ, Cox TM (2001) Tartrate-resistant acid phosphatase (Acp 5): identification in diverse human tissues and dendritic cells. J Histochem Cytochem 49:675–684PubMedGoogle Scholar
  6. 6.
    Ljusberg J, Ek-Rylander B, Andersson G (1999) Tartrate-resistant purple acid phosphatase is synthesized as a latent proenzyme and activated by cysteine proteinases. Biochem J 343 Pt 1:63-69Google Scholar
  7. 7.
    Ljusberg J, Wang Y, Lang P, Norgard M, Dodds R, Hultenby K, Ek-Rylander B, Andersson G (2005) Proteolytic excision of a repressive loop domain in tartrate-resistant acid phosphatase by cathepsin K in osteoclasts. J Biol Chem 280:28370–28381CrossRefPubMedGoogle Scholar
  8. 8.
    Oddie GW, Schenk G, Angel NZ, Walsh N, Guddat LW, de Jersey J, Cassady AI, Hamilton SE, Hume DA (2000) Structure, function, and regulation of tartrate-resistant acid phosphatase. Bone 27:575–584CrossRefPubMedGoogle Scholar
  9. 9.
    Janckila AJ, Takahashi K, Sun SZ, Yam LT (2001) Tartrate-resistant acid phosphatase isoform 5b as serum marker for osteoclastic activity. Clin Chem 47:74–80PubMedGoogle Scholar
  10. 10.
    Rissanen JP, Suominen MI, Peng Z, Halleen JM (2008) Secreted tartrate-resistant acid phosphatase 5b is a marker of osteoclast number in human osteoclast cultures and the rat ovariectomy model. Calcif Tissue Int 82:108–115CrossRefPubMedGoogle Scholar
  11. 11.
    Halleen JM, Ylipahkala H, Alatalo SL, Janckila AJ, Heikkinen JE, Suominen H, Cheng S, Vaananen HK (2002) Serum tartrate-resistant acid phosphatase 5b, but not 5a, correlates with other markers of bone turnover and bone mineral density. Calcif Tissue Int 71:20–25CrossRefPubMedGoogle Scholar
  12. 12.
    Halleen JM, Alatalo SL, Suominen H, Cheng S, Janckila AJ, Vaananen HK (2000) Tartrate-resistant acid phosphatase 5b: a novel serum marker of bone resorption. J Bone Miner Res 15:1337–1345CrossRefPubMedGoogle Scholar
  13. 13.
    Halleen JM, Alatalo SL, Janckila AJ, Woitge HW, Seibel MJ, Vaananen HK (2001) Serum tartrate-resistant acid phosphatase 5b is a specific and sensitive marker of bone resorption. Clin Chem 47:597–600PubMedGoogle Scholar
  14. 14.
    Adams LM, Warburton MJ, Hayman AR (2007) Human breast cancer cell lines and tissues express tartrate-resistant acid phosphatase (TRAP). Cell Biol Int 31:191–195CrossRefPubMedGoogle Scholar
  15. 15.
    Honig A, Rieger L, Kapp M, Krockenberger M, Eck M, Dietl J, Kammerer U (2006) Increased tartrate-resistant acid phosphatase (TRAP) expression in malignant breast, ovarian and melanoma tissue: an investigational study. BMC Cancer 6:199CrossRefPubMedGoogle Scholar
  16. 16.
    Chao TY, Yu JC, Ku CH, Chen MM, Lee SH, Janckila AJ, Yam LT (2005) Tartrate-resistant acid phosphatase 5b is a useful serum marker for extensive bone metastasis in breast cancer patients. Clin Cancer Res 11:544–550PubMedGoogle Scholar
  17. 17.
    Jung K, Lein M, Stephan C, Von Hosslin K, Semjonow A, Sinha P, Loening SA, Schnorr D (2004) Comparison of 10 serum bone turnover markers in prostate carcinoma patients with bone metastatic spread: diagnostic and prognostic implications. Int J Cancer 111:783–791CrossRefPubMedGoogle Scholar
  18. 18.
    Lyubimova NV, Pashkov MV, Tyulyandin SA, Gol’dberg VE, Kushlinskii NE (2004) Tartrate-resistant acid phosphatase as a marker of bone metastases in patients with breast cancer and prostate cancer. Bull Exp Biol Med 138:77–79PubMedGoogle Scholar
  19. 19.
    Mose S, Menzel C, Kurth AA, Obert K, Ramm U, Eberlein K, Boettcher HD, Pichlmeier U (2005) Evaluation of tartrate-resistant acid phosphatase (TRACP) 5b as bone resorption marker in irradiated bone metastases. Anticancer Res 25:4639–4645PubMedGoogle Scholar
  20. 20.
    Chung YC, Ku CH, Chao TY, Yu JC, Chen MM, Lee SH (2006) Tartrate-resistant acid phosphatase 5b activity is a useful bone marker for monitoring bone metastases in breast cancer patients after treatment. Cancer Epidemiol Biomarkers Prev 15:424–428CrossRefPubMedGoogle Scholar
  21. 21.
    Salminen E, Ala-Houhala M, Korpela J, Varpula M, Tiitinen SL, Halleen JM, Vaananen HK (2005) Serum tartrate-resistant acid phosphatase 5b (TRACP 5b) as a marker of skeletal changes in prostate cancer. Acta Oncol 44:742–747CrossRefPubMedGoogle Scholar
  22. 22.
    Ek-Rylander B, Barkhem T, Ljusberg J, Ohman L, Andersson KK, Andersson G (1997) Comparative studies of rat recombinant purple acid phosphatase and bone tartrate-resistant acid phosphatase. Biochem J 321(Pt 2):305–311PubMedGoogle Scholar
  23. 23.
    Lang P, Andersson G (2005) Differential expression of monomeric and proteolytically processed forms of tartrate-resistant acid phosphatase in rat tissues. Cell Mol Life Sci 62:905–918CrossRefPubMedGoogle Scholar
  24. 24.
    Wang Y, Norgard M, Andersson G (2005) N-glycosylation influences the latency and catalytic properties of mammalian purple acid phosphatase. Arch Biochem Biophys 435:147–156CrossRefPubMedGoogle Scholar
  25. 25.
    Zenger S, Hollberg K, Ljusberg J, Norgard M, Ek-Rylander B, Kiviranta R, Andersson G (2007) Proteolytic processing and polarized secretion of tartrate-resistant acid phosphatase is altered in a subpopulation of metaphyseal osteoclasts in cathepsin K-deficient mice. Bone 41:820–832CrossRefPubMedGoogle Scholar
  26. 26.
    Igarashi Y, Lee MY, Matsuzaki S (2001) Heparin column analysis of serum type 5 tartrate-resistant acid phosphatase isoforms. J Chromatogr B Biomed Sci Appl 757:269–276CrossRefPubMedGoogle Scholar
  27. 27.
    Wang Y, Andersson G (2007) Expression and proteolytic processing of mammalian purple acid phosphatase in CHO-K1 cells. Arch Biochem Biophys 461:85–94CrossRefPubMedGoogle Scholar
  28. 28.
    Zenger S, Ek-Rylander B, Andersson G (2010) Biogenesis of tartrate-resistant acid phosphatase isoforms 5a and 5b in stably transfected MDA-MB-231 breast cancer epithelial cells. Biochim Biophys Acta 1803:598–607CrossRefPubMedGoogle Scholar
  29. 29.
    Yam LT, Li CY, Lam KW (1971) Tartrate-resistant acid phosphatase isoenzyme in the reticulum cells of leukemic reticuloendotheliosis. N Engl J Med 284:357–360CrossRefPubMedGoogle Scholar
  30. 30.
    Mose S, Menzel C, Kurth AA, Obert K, Breidert I, Borowsky K, Bottcher HD (2003) Tartrate-resistant acid phosphatase 5b as serum marker of bone metabolism in cancer patients. Anticancer Res 23:2783–2788PubMedGoogle Scholar
  31. 31.
    Terpos E, de la Fuente J, Szydlo R, Hatjiharissi E, Viniou N, Meletis J, Yataganas X, Goldman JM, Rahemtulla A (2003) Tartrate-resistant acid phosphatase isoform 5b: a novel serum marker for monitoring bone disease in multiple myeloma. Int J Cancer 106:455–457CrossRefPubMedGoogle Scholar
  32. 32.
    Janckila AJ, Parthasarathy RN, Parthasarathy LK, Seelan RS, Hsueh YC, Rissanen J, Alatalo SL, Halleen JM, Yam LT (2005) Properties and expression of human tartrate-resistant acid phosphatase isoform 5a by monocyte-derived cells. J Leukoc Biol 77:209–218CrossRefPubMedGoogle Scholar
  33. 33.
    Venables JP (2004) Aberrant and alternative splicing in cancer. Cancer Res 64:7647–7654CrossRefPubMedGoogle Scholar
  34. 34.
    Ek-Rylander B, Bill P, Norgard M, Nilsson S, Andersson G (1991) Cloning, sequence, and developmental expression of a type 5, tartrate-resistant, acid phosphatase of rat bone. J Biol Chem 266:24684–24689PubMedGoogle Scholar
  35. 35.
    Baumbach GA, Saunders PT, Ketcham CM, Bazer FW, Roberts RM (1991) Uteroferrin contains complex and high mannose-type oligosaccharides when synthesized in vitro. Mol Cell Biochem 105:107–117CrossRefPubMedGoogle Scholar
  36. 36.
    Saunders PT, Renegar RH, Raub TJ, Baumbach GA, Atkinson PH, Bazer FW, Roberts RM (1985) The carbohydrate structure of porcine uteroferrin and the role of the high mannose chains in promoting uptake by the reticuloendothelial cells of the fetal liver. J Biol Chem 260:3658–3665PubMedGoogle Scholar
  37. 37.
    Dube DH, Bertozzi CR (2005) Glycans in cancer and inflammation–potential for therapeutics and diagnostics. Nat Rev Drug Discov 4:477–488CrossRefPubMedGoogle Scholar
  38. 38.
    Pollard JW (2009) Trophic macrophages in development and disease. Nat Rev Immunol 9:259–270CrossRefPubMedGoogle Scholar
  39. 39.
    Smyth MJ, Crowe NY, Godfrey DI (2001) NK cells and NKT cells collaborate in host protection from methylcholanthrene-induced fibrosarcoma. Int Immunol 13:459–463CrossRefPubMedGoogle Scholar
  40. 40.
    Hayakawa Y, Takeda K, Yagita H, Smyth MJ, Van Kaer L, Okumura K, Saiki I (2002) IFN-gamma-mediated inhibition of tumor angiogenesis by natural killer T-cell ligand, alpha-galactosylceramide. Blood 100:1728–1733PubMedGoogle Scholar
  41. 41.
    Bune AJ, Hayman AR, Evans MJ, Cox TM (2001) Mice lacking tartrate-resistant acid phosphatase (Acp 5) have disordered macrophage inflammatory responses and reduced clearance of the pathogen, Staphylococcus aureus. Immunology 102:103–113CrossRefPubMedGoogle Scholar
  42. 42.
    Kong YY, Yoshida H, Sarosi I, Tan HL, Timms E, Capparelli C, Morony S, Oliveira-dos-Santos AJ, Van G, Itie A, Khoo W, Wakeham A, Dunstan CR, Lacey DL, Mak TW, Boyle WJ, Penninger JM (1999) OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature 397:315–323CrossRefPubMedGoogle Scholar
  43. 43.
    Wittrant Y, Theoleyre S, Couillaud S, Dunstan C, Heymann D, Redini F (2003) Regulation of osteoclast protease expression by RANKL. Biochem Biophys Res Commun 310:774–778CrossRefPubMedGoogle Scholar
  44. 44.
    Jones DH, Nakashima T, Sanchez OH, Kozieradzki I, Komarova SV, Sarosi I, Morony S, Rubin E, Sarao R, Hojilla CV, Komnenovic V, Kong YY, Schreiber M, Dixon SJ, Sims SM, Khokha R, Wada T, Penninger JM (2006) Regulation of cancer cell migration and bone metastasis by RANKL. Nature 440:692–696CrossRefPubMedGoogle Scholar
  45. 45.
    Simpson KJ, Selfors LM, Bui J, Reynolds A, Leake D, Khvorova A, Brugge JS (2008) Identification of genes that regulate epithelial cell migration using an siRNA screening approach. Nat Cell Biol 10:1027–1038CrossRefPubMedGoogle Scholar
  46. 46.
    Ek-Rylander B, Andersson G (2010) Osteoclast migration on phosphorylated osteopontin is regulated by endogenous tartrate-resistant acid phosphatase. Exp Cell Res 316:443–451CrossRefPubMedGoogle Scholar
  47. 47.
    Sung V, Gilles C, Murray A, Clarke R, Aaron AD, Azumi N, Thompson EW (1998) The LCC15-MB human breast cancer cell line expresses osteopontin and exhibits an invasive and metastatic phenotype. Exp Cell Res 241:273–284CrossRefPubMedGoogle Scholar
  48. 48.
    Weber GF, Ashkar S, Cantor H (1997) Interaction between CD44 and osteopontin as a potential basis for metastasis formation. Proc Assoc Am Physicians 109:1–9PubMedGoogle Scholar
  49. 49.
    Christensen B, Kazanecki CC, Petersen TE, Rittling SR, Denhardt DT, Sorensen ES (2007) Cell type-specific post-translational modifications of mouse osteopontin are associated with different adhesive properties. J Biol Chem 282:19463–19472CrossRefPubMedGoogle Scholar
  50. 50.
    Weber GF, Zawaideh S, Hikita S, Kumar VA, Cantor H, Ashkar S (2002) Phosphorylation-dependent interaction of osteopontin with its receptors regulates macrophage migration and activation. J Leukoc Biol 72:752–761PubMedGoogle Scholar
  51. 51.
    Sun P, Sleat DE, Lecocq M, Hayman AR, Jadot M, Lobel P (2008) Acid phosphatase 5 is responsible for removing the mannose 6-phosphate recognition marker from lysosomal proteins. Proc Natl Acad Sci USA 105:16590–16595CrossRefPubMedGoogle Scholar
  52. 52.
    Hayman AR, Cox TM (1994) Purple acid phosphatase of the human macrophage and osteoclast. Characterization, molecular properties, and crystallization of the recombinant di-iron-oxo protein secreted by baculovirus-infected insect cells. J Biol Chem 269:1294–1300PubMedGoogle Scholar
  53. 53.
    Nishikawa M (2008) Reactive oxygen species in tumor metastasis. Cancer Lett 266:53–59CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Serhan Zenger
    • 1
  • Wentao He
    • 1
  • Barbro Ek-Rylander
    • 1
  • Daphne Vassiliou
    • 1
  • Rickard Wedin
    • 2
  • Henrik Bauer
    • 2
  • Göran Andersson
    • 1
  1. 1.Division of Pathology F 46, Department of Laboratory MedicineKarolinska Institute, Karolinska University HospitalHuddingeSweden
  2. 2.Oncology Service, Department of OrthopaedicsKarolinska University HospitalStockholmSweden

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