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. 1997 Feb;123(2):71–81. doi: 10.1007/BF01269884

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

Geoffrey M Curtin 1,✉, Susan M Fischer 1, Thomas J Slaga 1
PMCID: PMC12201151  PMID: 9030245

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.

Key Words: Autocrine regulation, Progression factor, Skin epithelium

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

Footnotes

This work was supported by NIH grant CA-57596

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