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AGE-R3/galectin-3 expression in osteoblast-like cells: Regulation by AGEs

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Abstract

The accumulation of irreversible advanced glycation endproducts (AGEs) on long-lived proteins, and the interaction of AGEs with cellular receptors such as AGE-R3/galectin-3 and RAGE, are considered to be key events in the development of long-term complications of diabetes mellitus, Alzheimer's disease, uremia and ageing. The aim of this study was to investigate the expression and sub-cellular distribution of galectin-3, as well as its possible modulation by AGEs, in MC3T3E1 mouse calvaria-derived osteoblasts and in UMR 106 rat osteosarcoma cells. Both osteoblastic lines were cultured either with control bovine serum albumin (BSA) or with AGEs-BSA for 48 h. Cells were evaluated for galectin-3 expression by fixing and immunofluorescent microscopic analysis; or Western blot analysis of whole cell extracts, sub-cellular fractions and culture media. Both cell lines express 30 kDa (monomeric) galectin-3, although expression was about 15-fold lower in the UMR106 osteosarcoma cells. Dimeric (70 kDa) galectin-3 was additionally observed in the UMR106 cells. Immunofluorescent analysis of galectin-3 distribution showed a diffuse cytoplasmic and strong nuclear pattern in MC3T3E1 osteoblasts, and a patchy cytoplasmic pattern in UMR106 cells. Western blot analysis for both cell lines showed that galectin-3 was mainly found in the cytoplasm and in minor amounts in the microsomal fraction, while considerable amounts were secreted into the culture media. Exposure to 100–200 μg/mL AGEs-BSA increased the cellular content of 30 kDa galectin-3 (20–25% for MC3T3E1 and 35–70% for UMR106 versus control BSA, p < 0.05), and decreased the culture media levels of galectin-3 (10–20% for MC3T3E1 and for UMR106 versus control BSA, p < 0.05). These results confirm the expression of galectin-3 in osteoblastic cells, and suggest different levels and sub-cellular distribution of this protein in transformed versus non-transformed osteoblasts. Osteoblastic exposure to AGEs alters their expression and secretion of galectin-3, which could have significant consequences on osteoblast metabolism and thus on bone turnover (Mol Cell Biochem 266: 17–24, 2004)

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References

  1. Vlassara H, Bucala R, Striker L: Pathogenic effects of advanced glycosylation: Biochemical, biologic and clinical implications for diabetes and ageing. Lab Invest 70: 138–151, 1994

    Google Scholar 

  2. Baynes JW, Thorpe SR: Role of oxidative stress in diabetic complications: A new perspective on an old paradigm. Diabetes 48: 1–9, 1999

    Google Scholar 

  3. Thomalley PJ: Cell activation by glycated proteins. Age receptors, receptor recognition factors and functional classification of AGEs. Cell Mol Biol 44: 1013–1033, 1998

    Google Scholar 

  4. McCarthy AD, Etcheverry SB, Bruzzone L, Cortizo AM: Effects of advanced glycation end-products on the proliferation and differentiation of osteoblast-like cells. Mol Cell Biochem 170: 43–51, 1997

    Google Scholar 

  5. McCarthy AD, Etcheverry SB, Cortizo AM: Advanced glycation end product-specific receptors in rat and mouse osteoblast-like cells: Regulation with stages of differentiation. Acta Diabetol 36: 45–52, 1999

    Google Scholar 

  6. Schmidt AM, Yan SD, Yan SF, Stem DM: The biology of the receptor for advanced glycation end products and its ligands. Biochim Biophys Acta 1498: 99–111, 2000

    Google Scholar 

  7. Cortizo AM, Lettieri M, Barrio DA, Mercer N, Etcheverry S, McCarthy AD: Advanced glycation end-products (AGEs) induce concerted changes in the osteoblastic expression of their receptor RAGE and in the activation of extracellular signal-regulated kinases (ERK). Mol Cell Biochem 250: 1–10, 2003

    Google Scholar 

  8. Barontes SH, Cooper DNW, Gitt MA, Leffler H: Galectins. Structure and function of a large family of animal lectins. J Biol Chem 269: 20807–20810, 1994

    Google Scholar 

  9. Liu F-T, Patterson RJ, Wang JL: Intracellular functions of galectins. Biochim Biophys Acta 1572: 263–273, 2002

    Google Scholar 

  10. Wang L, Inohara H, Pienta KJ, Raz A: Galectin-3 is a nuclear matrix protein which binds RNA. Biochem Biophys Res Commun 217: 292–303, 1995

    Google Scholar 

  11. Vlassara H, Li YM, Imani F, Wojciechowicz D, Yang Z, Liu FT, Cerami A: Identification of galectin-3 as a high-affinity binding protein for advanced glycation end products (AGE): A new member of the AGE-receptor complex. Mol Med 1: 634–646, 1995

    Google Scholar 

  12. Levin ME, Boisseau BC, Avioli LVL: Effects of diabetes mellitus on bone mass in juvenile and adult-onset diabetes. New Engl J Med 294: 241–245, 1976

    Google Scholar 

  13. Krakauer JC, McKenna MJ, Buderer NF, Rao DS, Whitehouse FW, Parfitt AM. Bone loss and bone turnover in diabetes mellitus. Diabetes 44: 775–782, 1995

    Google Scholar 

  14. Laemmli UK: Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970

    Google Scholar 

  15. Lowry OH, Rosebrough NJ, Farr AJ, Randall RJ: Protein measurement with Folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  16. Sálice VC, Cortizo AM, Gómez Dumm CL, Etcheverry SB: Tyrosine phosphorylation and morphological transformation induced by four vanadium compounds on MC3T3E1 cells. Mol Cell Biochem 198: 119–128, 1999

    Google Scholar 

  17. McCarthy AD, Etcheverry SB, Bruzzone L, Lettieri MG, Barrio DA, Cortizo AD. Non-enzymatic glycosylation of a type I collagen matrix: Effects on osteoblastic development and oxidative stress. BMC Cell Biol 2: 16, 2001

    Google Scholar 

  18. Vlassara H, Brownlee M, Cerami A: High-affinity receptor-mediated uptake and degradation of glucose-modified proteins; a potential mechanism for the removal of senescent macromolecules. Proc Natl Acad Sci USA 82: 5588–5592, 1985

    Google Scholar 

  19. Yang Z, Makita Z, Horii Y, Brunelle S, Cerami A, Sehajpal P, Suthanthiran M, Vlassara H: Two novel rat liver membrane proteins that bind advanced glycosylation endproducts: Relationship to macrophage receptor for glucose-modified proteins. J Exp Med 174: 515–524, 1991

    Google Scholar 

  20. Tomasek JJ, Meyers SW, Basinger JB, Green DJ, Shew RL. Diabetic and age-related enhancement of collagen-linked fluorescence in cortical bones of rats. Life Sci 55: 855–861, 1994

    Google Scholar 

  21. Katayama Y, Akatsu T, Yamamoto M, Kugai N, Nagata N: Role of nonenzymatic glycosylation of type I collagen in diabetic osteopenia. J Bone Miner Res 11: 931–937, 1996

    Google Scholar 

  22. Aubin JE, Gupta AK, Bhargava U, Turksen K: Expression and regulation of galectin 3 in rat osteoblastic cells. J Cell Physiol 169: 468–480, 1996

    Google Scholar 

  23. Quarles LD, Yahay DA, Lever LW, Caton R, Wenstrup RJ: Distinct proliferative and differentiated stages of murine MC3T3E1 cells in culture: An in vitro model of osteoblast development. J Bone Miner Res 7: 683–692, 1992

    Google Scholar 

  24. Partridge NC, Alcorn D, Michelangeli VP, Ryan G, Martin TJ: Morphological and biochemical characterization of four clonal osteogenic sarcoma cell lines of rat origin. Cancer Res: 43: 4308–4312, 1983

    Google Scholar 

  25. Openo KP, Kadrofske MM, Patterson RJ, Wang JL: Galectin-3 expression and subcellular localization in senescent human fibroblasts. Exp Cell Res 255: 2778–290, 2000

    Google Scholar 

  26. van den Brule FA, Waltregny D, Liu FT, Castronovo V: Alteration of the cytoplasmic/nuclear expression pattern of galectin-3 correlates with prostate carcinoma progression. Int J Cancer 89: 361–367, 2000

    Google Scholar 

  27. Honjo Y, Inohara H, Akahani S, Yoshii T, Takenaka Y, Yoshida J, Hattori K, Tomiyama Y, Raz A, Kubo T: Expression of cytoplasmic galectin-3 as a prognostic marker in tongue carcinoma. Clin Cancer Res 6: 4635–4640, 2000

    Google Scholar 

  28. Yu F, Finley RL, Raz A, Kim HRC: Galectin-3 translocates to the perinuclear membranes and inhibits cytochrome c release from the mitochondria. A role for synexin in galectin-3 translocation. J Biol Chem 277: 15819–15827, 2002

    Google Scholar 

  29. Stock M, Schäfer H, Stricker S, Gross G, Mundlos S, Otto F: Expression of galectin-3 in skeletal tissues is controlled by Runx2. J Biol Chem 278: 17360–17367, 2003

    Google Scholar 

  30. Pugliese G, Pricci F, Leto G, Amadio L, Iacobini C, Romeo G, Lenti L, Sale P, Gradini R, Liu F, Di Mario U: The diabetic milieu modulates the advanced glycation end product-receptor complex in the mesangium by inducing or upregulating galectin-3 expression. Diabetes 49: 1249–1257, 2000

    Google Scholar 

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Authors and Affiliations

  1. Cátedra de Bioquímica Patológica, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina

    Natalia Mercer, Antonio D. McCarthy, Susana B. Etcheverry & Ana M. Cortizo

  2. Center of Marine Biotechnology, University of Maryland, Baltimore, MD, 21202-3101, USA

    Hafiz Ahmed & Gerardo R. Vasta

Authors
  1. Natalia Mercer
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  2. Hafiz Ahmed
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  3. Antonio D. McCarthy
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  4. Susana B. Etcheverry
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  5. Gerardo R. Vasta
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  6. Ana M. Cortizo
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Mercer, N., Ahmed, H., McCarthy, A.D. et al. AGE-R3/galectin-3 expression in osteoblast-like cells: Regulation by AGEs. Mol Cell Biochem 266, 17–24 (2004). https://doi.org/10.1023/B:MCBI.0000049128.71095.ac

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