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Identification of an autocrine mechanism for regulating cell-cycle progression in murine keratinocytes

  • Original Paper
  • Experimental Oncology
  • Published:
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Abstract

Primary murine keratinocytes possess a limited doubling potential regardless of plating density or the inclusion of competence factors insulin, epidermal growth factor, and/or fetal bovine serum within the culture medium. In contrast, a murine cell line (CH-72), derived from a 7,12-dimethylbenz[a]-anthraceneinitiated, 12-O-tetra-decanoylphorbol-13-acetate-promoted mouse skin carcinoma, was found to exhibit unlimited proliferative potential; this was demonstrated by the ability of these cells to produce the progression factor required for entry into the DNA-synthesis phase in the absence of competence-factor stimulation. Conditioned medium, collected from murine carcinoma cells, was subsequently shown to increase the level of [3H] thymidine incorporation in competence-factor-deprived CH-72 cultures by more than a factor of 4 within 16h. Moreover, consistent with its ability of recruit cells cycling within the first gap phase directly into the DNA-synthesis phase, the autocrine progression factor present in conditioned medium decreased the G1 ∶ S ratio from the 55 ∶ 29 observed with growth medium controls to 38 ∶ 46. Preliminary characterization of the autocrine factor produced by cultured murine carcinoma cells using gel-filtration chromatography revealed a molecular mass of less than 2 kDa, similar in size to the factor previously shown by our laboratory to promote G1-phase progression in cultures of normal human foreskin keratinocytes.

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Abbreviations

PDGF :

platelet-derived growth factor

IGF :

insulin-like growth factor

IL-2 :

interleukin-2

PBS :

phosphate-buffered saline

EMEM :

Eagle's mininal essential medium

EGF :

epidermal growth factor

FBS :

fetal bovine serum

CDK :

cyclin-dependent kinase

TGF :

transforming growth factor

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Authors and Affiliations

  1. Department of Carcinogenesis, University of Texas M.D. Anderson Cancer Center-Science Park Research Division, P. O. Box 389, 78957, Smithville, TX, USA

    Geoffrey M. Curtin, Susan M. Fischer & Thomas J. Slaga

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  1. Geoffrey M. Curtin
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  2. Susan M. Fischer
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  3. Thomas J. Slaga
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This work was supported by NIH grant CA-57596

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Curtin, G.M., Fischer, S.M. & Slaga, T.J. Identification of an autocrine mechanism for regulating cell-cycle progression in murine keratinocytes. J Cancer Res Clin Oncol 123, 71–81 (1997). https://doi.org/10.1007/BF01269884

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  • DOI: https://doi.org/10.1007/BF01269884

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