Role of p14^ARF in Replicative and Induced Senescence of Human Fibroblasts

Abstract

Following a proliferative phase of variable duration, most normal somatic cells enter a growth arrest state known as replicative senescence. In addition to telomere shortening, a variety of environmental insults and signaling imbalances can elicit phenotypes closely resembling senescence. We used p53^−/− and p21^−/− human fibroblast cell strains constructed by gene targeting to investigate the involvement of the Arf-Mdm2-p53-p21 pathway in natural as well as premature senescence states. We propose that in cell types that upregulate p21 during replicative exhaustion, such as normal human fibroblasts, p53, p21, and Rb act sequentially and constitute the major pathway for establishing growth arrest and that the telomere-initiated signal enters this pathway at the level of p53. Our results also revealed a number of significant differences between human and rodent fibroblasts in the regulation of senescence pathways.

ACKNOWLEDGMENTS

We gratefully acknowledge J. Morgenstern for pWZL-Blast retrovirus vector, S. Lowe for Ha-(G12V)Ras cDNA, D. Sidransky for a P1 clone of the genomic INK4a locus, R. Weinberg for hTERT cDNA, Y. Xiong for Arf cDNA, and D. DiMaio for helpful comments on the manuscript.

This work was supported by NIH grant AG16694 to J.M.S.