beta-catenin signalling modulates proliferative potential of human epidermal keratinocytes independently of intercellular adhesion.
We found that cultured human keratinocytes with high proliferative potential, the putative epidermal stem cells, expressed a higher level of noncadherin-associated beta-catenin than populations enriched for keratinocytes of lower proliferative potential. To investigate the physiological significance of this, a series of beta-catenin constructs was introduced into keratinocytes via retroviral infection. Full-length beta-catenin and a mutant containing only nine armadillo repeats had little effect on proliferative potential in culture, the full-length protein being rapidly degraded. However, expression of stabilised, N-terminally truncated beta-catenin increased the proportion of putative stem cells to almost 90% of the proliferative population in vitro without inducing malignant transformation, and relieved the differentiation stimulatory effect of overexpressing the E-cadherin cytoplasmic domain. Conversely, beta-catenin lacking armadillo repeats acted as a dominant negative mutant and stimulated exit from the stem cell compartment in culture. The positive and negative effects of the beta-catenin mutants on proliferative potential were independent of effects on cell-cycle kinetics, overt terminal differentiation or intercellular adhesion, and correlated with stimulation or inhibition of transactivation of a TCF/LEF reporter in basal keratinocytes. We conclude that the elevated level of cytoplasmic beta-catenin in those keratinocytes with characteristics of epidermal stem cells contributes to their high proliferative potential.
Zhu
A J
AJ
Keratinocyte Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, England.
Watt
F M
FM
D000818:Animals; D015820:Cadherins / Q000235:genetics; D002448:Cell Adhesion; D002453:Cell Cycle; D002454:Cell Differentiation; D002455:Cell Division; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D000078404:Epidermal Cells; D015870:Gene Expression; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000502:physiology; D016296:Mutagenesis; D015398:Signal Transduction; D013234:Stem Cells; D015534:Trans-Activators; D015533:Transcriptional Activation; D014981:Xenopus; D029867:Xenopus Proteins; D051176:beta Catenin
C495266:CTNNB1 protein, Xenopus; C495270:CTNNB1 protein, human; D015820:Cadherins; D003598:Cytoskeletal Proteins; D015534:Trans-Activators; D029867:Xenopus Proteins; D051176:beta Catenin