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Basic Fibroblast Growth Factor Autocrine Loop Controls Human Osteosarcoma Phenotyping and Differentiation

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Abstract

Background

We focused on the phenotype of non-mineralizing MG63 and mineralizing TE85 human osteosarcoma cells and investigated the role of bFGF in modulating their differentiative responses. Basic FGF expression and bFGF effects on osteocalcin, runt-related transcription factor-2 (RUNX2), matrix molecular production and bFGF receptors, were evaluated.

Materials and Methods

Osteocalcin and RUNX2 gene expression were studied by RT-PCR analysis. We evaluated cell proliferation by DNA content and performed differentiation studies on glycosaminoglican (GAG), collagen and proteoglican (PG) synthesis by using radiolabelled precursors and Northern blotting. BFGF receptors were quantified by bFGF receptor binding assay.

Results

Osteocalcin is expressed in MG63 and TE65. RUNX2 RNA is differentially spliced in the two cell lines. BFGF elicits the effects of differentially splicing RUNX2. Proliferation, GAG synthesis, bFGF and proteoglycan mRNA expression, high and low affinity bFGF receptors, were more marked in MG63 and differently affected by bFGF. Procollagen expression and alkaline phosphatase activity were significantly reduced. BFGF increased TE85 cell proliferation and reduced TE85 procollagen and osteocalcin production.

Conclusions

The different splice variants in RUNX2 gene in the two cell lines might be related to their different phenotypes. The less differentiated stage of MG63 could also be related to bFGF over-production and more bFGF receptors. The consequent increase in bFGF-bFGF receptor binding could explain the bFGF differentiative effects on MG63. We suggest an autocrine role of bFGF endogenous release in controlling the different osteosarcoma phenotypes.

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Acknowledgments

The work was supported by a grant from C.N.R., Ministero Università Ricerca Scientifica Tecnologica (M.I.U.R.) and Fondazione Cassa di Risparmio di Perugia (SA 172901 project). We are indebted to Dr. Larry W. Fisher (National Institutes of Health, Bethesda, MD, USA) for biglycan and decorin core protein cDNAs; to Dr. Judith Abraham, PhD (Scios Nova, CA, USA) for bFGF cDNA; to Dr. Eero Vuorio (University of Turku, Finland) for procollagen α1 (I) cDNA; to Dr. M. Bernfield (Newborn Medicine, Children’s Hospital, Boston, MA, USA) for syndecan-1 cDNA.

We thank Dr. Geraldine Boyd for English language editing.

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Authors and Affiliations

  1. Sezione di Istologia, Facoltà di Medicina, Università degli Studi di Perugia, Via del Giochetto 06126, Perugia, Italy

    Maria Bodo, Cinzia Lilli, Catia Bellucci, Mario Calvitti, Silvia Bellocchio, Chiara Balducci & Tiziano Baroni

  2. Istituto di Istologia ed Embriologia generale, Università degli Studi di Bologna, Bologna, Italy

    Paolo Carinci & Furio Pezzetti

  3. Dipartimento di Anatomia Umana, Università Statale di Milano, Milano, Italy

    Giordano Stabellini

  4. Cattedra di Chirurgia Maxillo-Facciale, Università di Ferrara, Ferrara, Italy

    Francesco Carinci

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  1. Maria Bodo
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Correspondence to Maria Bodo.

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*These authors contributed equally to the project.

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Bodo, M., Lilli, C., Bellucci, C. et al. Basic Fibroblast Growth Factor Autocrine Loop Controls Human Osteosarcoma Phenotyping and Differentiation. Mol Med 8, 393–404 (2002). https://doi.org/10.1007/BF03402020

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