Elsevier

Experimental Gerontology

Volume 35, Issue 8, October 2000, Pages 927-945
Experimental Gerontology

Perspective
Cellular and molecular mechanisms of stress-induced premature senescence (SIPS) of human diploid fibroblasts and melanocytes

https://doi.org/10.1016/S0531-5565(00)00180-7Get rights and content

Abstract

Replicative senescence of human diploid fibroblasts (HDFs) or melanocytes is caused by the exhaustion of their proliferative potential. Stress-induced premature senescence (SIPS) occurs after many different sublethal stresses including H2O2, hyperoxia, or tert-butylhydroperoxide. Cells in replicative senescence share common features with cells in SIPS: morphology, senescence-associated β-galactosidase activity, cell cycle regulation, gene expression and telomere shortening. Telomere shortening is attributed to the accumulation of DNA single-strand breaks induced by oxidative damage. SIPS could be a mechanism of accumulation of senescent-like cells in vivo. Melanocytes exposed to sublethal doses of UVB undergo SIPS. Melanocytes from dark- and light- skinned populations display differences in their cell cycle regulation. Delayed SIPS occurs in melanocytes from light-skinned populations since a reduced association of p16Ink-4a with CDK4 and reduced phosphorylation of the retinoblastoma protein are observed. The role of reactive oxygen species in melanocyte SIPS is unclear. Both replicative senescence and SIPS are dependent on two major pathways. One is triggered by DNA damage, telomere damage and/or shortening and involves the activation of the p53 and p21waf-1 proteins. The second pathway results in the accumulation of p16Ink-4a with the MAP kinase signalling pathway as possible intermediate. These data corroborate the thermodynamical theory of ageing, according to which the exposure of cells to sublethal stresses of various natures can trigger SIPS, with possible modulations of this process by bioenergetics.

Section snippets

Free radicals, ageing, and replicative senescence

The free radical theory of ageing (Harman et al., 1998) proposes that normal ageing results from random deleterious damage to tissue by oxidative stress caused by reactive oxygen species (ROS). Aged animals contain defective mitochondria and can produce higher levels of ROS than their young counterparts. Ischemia-reperfusion and inflammation produce ROS, as well as many xenobiotics do. Tissues from aged individuals or aged experimental animals accumulate oxidative damage in their DNA, proteins

Induction of premature senescence by oxidative stress

From an experimental point of view, oxidative stress is probably the most often used inducer of SIPS. Common pathways with other triggers will be discussed below. In accordance with theoretical studies based on the criteria of stability of cellular systems (Toussaint et al., 1991), numerous experimental studies demonstrated that sublethal oxidative stresses induce premature replicative senescence. Both chronic and acute oxidative stress protocols were used to induce cellular senescence. In the

Morphological studies

HDFs exposed to repeated sublethal stress under t-BHP display the morphological phenotype of senescence. The first studies were based on the description of the successive HDF morphotypes observed during in vitro ageing: three successive mitotic morphotypes followed by three successive post-mitotic morphotypes and eventually a degenerative state of short existence, all of these states observed at very low density allowing spreading of cells (Rodemann et al., 1989, Toussaint et al., 1992). Using

Telomeres and SIPS

There are different causes of telomere shortening with replicative age. The inability of DNA polymerases to replicate a linear DNA template to its very end (the so-called end-replication problem) has been known for a long time (Olovnikov, 1973). The contribution of the end-replication problem to the actual telomere shortening might be as small as very few nucleotides per CPD, though. The action of a C strand-specific exonuclease has been suggested to contribute to the shortening of telomeres (

SIPS: interaction between metabolic and signalling pathways

Antioxidants are not the only ones which may slow down the rate of oxidative stress-induced premature senescence (SIPS). A thermodynamic model of ageing was proposed that predicts not only that sublethal stress may induce SIPS, but also that the level of global metabolic activity may protect the cells against SIPS and against cell death caused by lethal stress (Remacle et al., 1995, Toussaint et al., 1995a). Under stress, additional energy utilisation is required for the induction of defence

Premature senescence in human melanocytes

Melanocytes are pigment cells that are located in the basal layer of the epidermis. They produce the pigment melanin, which is synthesised in melanosomes, a lysosomal-like organelle. Melanin pigments play a key protective role against the carcinogenic effect of solar irradiation in vivo. In melanocytes, senescence is associated with increased binding of p16INK4a to CDK4 and loss of binding activity, and protein expression of transcription factors of the E2F family. Intriguingly, melanocytes

Conclusions

Ageing is the result of an complicated interplay of deterministic and stochastic processes. Random damage is prominent among the stochastic processes, leading to concepts like “critical threshold of error accumulation” (Remacle et al., 1992) or “failure of maintenance” (Holliday, 1988). These concepts encompass the notion that the kinetics of the accumulation of damage must not only be attributed to the extent of damage but also to the efficiency of the systems for elimination and/or repair of

Acknowledgements

O. Toussaint is a Research Assistant of the FNRS, Belgium. He also thanks the FRIA and the French Community of Belgium, the European Union Biomed & Health Research Programme, Shared-Cost Action Programme ‘Genage’ (BMH2 CT98) and the Fulbright programme. E.E. Medrano was supported by a grant AG3663 from the National Institute of Health (EEM). T. von Zglinicki thanks the Deutsche Forschungsgemeinschaft and the VERUM Foundation for support.

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