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IGF-I stimulates proliferation of spontaneously immortalized human keratinocytes (HACAT) by autocrine/paracrine mechanisms

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Abstract

HaCaT keratinocytes are derived from adult human skin and although spontaneously immortalized, remain highly related to their normal counterparts. We observed that HaCaT cells can proliferate in serum-free medium (SFM), in contrast to normal human keratinocytes whose growth in vitro requires a feeder layer and/or the supplementation with hormones and growth factors. Since autocrine production of growth factors has been proposed as the pathway that cells may exploit to escape growth regulation, we have investigated whether this is occurring in HaCaT cultured in SFM. Either epidermal growth factor (EGF) or insulin-like growth factor-1 (IGF-I) was effective and dose-dependently stimulated HaCaT replication. The ability of these keratinocytes to express EGF and IGF-I and their receptors was investigated by northern blot and reverse transcriptase-polymerase chain reaction (RT-PCR). We report that HaCaT cells synthesize mRNAs for IGF-I, IGF-II, IGF-IR and EGFR but not EGF mRNA. Immunoneutralization of IGF-I with specific monoclonal antibodies blocked spontaneous HaCaT proliferation in SFM, as did incubation with antibodies against IGF-IR. These data demonstrate that an autocrine/paracrine loop based on IGF-I may allow HaCaT keratinocytes to proliferate autonomously in culture in contrast to keratinocytes in primary culture. A similar mechanism may be involved in the development of hyperproliferative diseases of human skin and its functional disruption may represent the target for therapeutic approaches.

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Authors and Affiliations

  1. Medical and Surgical Outpatient Unit, Istituto Dermopatico dell’Immacolata, I.R.C.C.S., Saronno, Italy

    G. Pozzi

  2. Department of Pharmacology, University of Milan, Milan, Italy

    M. Guidi, F. Laudicina & E. E. Müller

  3. Regiona Reference Centre for Human Skin Culture, Niguarda Ca’ Granda Hospital, Milan, Italy

    M. Marazzi & L. Falcone

  4. Dermatologic Clinic IV, S. Paolo Hospital, University of Milan, Milan, Italy

    R. Betti & C. Crosti

  5. London Regional Cancer Center, Departments of Oncology and Biochemistry, University of Western Ontario, London, Ontario, Canada

    G. E. DiMattia

  6. Department of Experimental and Environmental Medicine and Biotechnology, University of Milano-Bicocca, via Cadore 48, 20052, Monza (Milano), Italy

    V. Locatelli & A. Torsello Ph.D.

Authors
  1. G. Pozzi
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  2. M. Guidi
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  3. F. Laudicina
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  4. M. Marazzi
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  5. L. Falcone
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  6. R. Betti
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  7. C. Crosti
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  8. E. E. Müller
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  9. G. E. DiMattia
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  10. V. Locatelli
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  11. A. Torsello Ph.D.
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Correspondence to A. Torsello Ph.D..

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Pozzi, G., Guidi, M., Laudicina, F. et al. IGF-I stimulates proliferation of spontaneously immortalized human keratinocytes (HACAT) by autocrine/paracrine mechanisms. J Endocrinol Invest 27, 142–149 (2004). https://doi.org/10.1007/BF03346259

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