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Cited by 103Telomeres, Telomerase, and SenescenceGenetics, Aging, and Longevity in Model OrganismsCancer-related Molecular Pathways

Normal Human Fibroblasts Are Resistant to RAS-Induced Senescence

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Benanti, Jennifer A., Galloway, Denise A.·2004·Molecular and Cellular Biology
DOI 10.1128/mcb.24.7.2842-2852.2004PMC PMC371123
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Abstract

Oncogenic stimuli are thought to induce senescence in normal cells in order to protect against transformation and to induce proliferation in cells with altered p53 and/or retinoblastoma (Rb) pathways. In human fibroblasts, RAS initiates senescence through upregulation of the cyclin-dependent kinase inhibitor p16INK4A. We show here that in contrast to cultured fibroblast strains, freshly isolated normal fibroblasts are resistant to RAS-induced senescence and instead show some characteristics of transformation. RAS did not induce growth arrest or expression of senescence-associated beta-galactosidase, and Rb remained hyperphosphorylated despite elevated levels of p16. Instead, RAS promoted anchorage-independent growth of normal fibroblasts, although expression of hTert with RAS increased colony formation and allowed normal fibroblasts to bypass contact inhibition. To test the hypothesis that p16 levels determine how cells respond to RAS, we expressed RAS in freshly isolated fibroblasts that expressed very low levels of p16, in hTert-immortalized fibroblasts that had accumulated intermediate levels of p16, and in IMR90 fibroblasts with high levels of p16. RAS induced growth arrest in cells with higher p16 levels, and this effect was reversed by p16 knockdown in the hTert-immortalized fibroblasts. These findings indicate that culture-imposed stress sensitizes cells to RAS-induced arrest, whereas early passage cells do not arrest in response to RAS.

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  • There appears to be a threshold level that renders human cells susceptible to RAS-induced senescence and beyond which human cells cannot proliferate.

    “Nonetheless, there appears to be a threshold level that renders human cells susceptible to RAS-induced senescence (Benanti and Galloway, 2004) and beyond which human cells cannot proliferate.”

    From Senescent cells, tumor suppression, and organismal aging: good citizens, bad neighbors (2005) · cites this paper as Benanti and Galloway, 2004

    The Results report that higher p16 levels sensitized human fibroblasts to RAS-induced senescence and that those cells arrested, while reducing p16 prevented arrest. This supports a threshold-like interpretation for the fibroblasts studied, but not the broader claim that human cells generally cannot proliferate beyond such a threshold. (LLM, conf=0.85)

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  • At least some cells express p16 in vivo, during long-term culture, and at elevated levels upon replicative senescence.

    “However, at least some cells express p16 in vivo (Krishnamurthy et al., 2004), during long-term culture (Benanti and Galloway, 2004), and at elevated levels upon replicative senescence (Alcorta et al., 1996; Hara et al., 1996; Brenner et al., 1998; Stein et al., 1999; Itahana et al., 2003).”

    From Senescent cells, tumor suppression, and organismal aging: good citizens, bad neighbors (2005) · cites this paper as Benanti and Galloway, 2004

  • Cell culture studies indicate that p16 prevents the reversal of senescence by p53 inactivation and is required for RAS-induced senescence.

    “Cell culture studies indicate that p16 prevents the reversal of senescence by p53 inactivation (Beausejour et al., 2003) and is required for RAS-induced senescence (Brookes et al., 2002; Benanti and Galloway, 2004).”

    From Senescent cells, tumor suppression, and organismal aging: good citizens, bad neighbors (2005) · cites this paper as Benanti and Galloway, 2004

  • Cells with low baseline p16 levels do not undergo OIS but show anchorage-independent growth, a feature of cell transformation.

    “Moreover, cells with low baseline p16 levels do not undergo OIS but show anchorage-independent growth (a feature of cell transformation) [54].”

    From Mechanisms and Regulation of Cellular Senescence (2021) · cites this paper as [54]

  • Culture of primary mouse and human fibroblasts at low physiological oxygen levels prevents senescence activation after long culturing or upon expression of activated Ras.

    “Culture of primary mouse and human fibroblasts at low physiological oxygen levels prevents senescence activation after long culturing (increasing passage number) or upon expression of activated Ras [54,111].”

    From Mechanisms and Regulation of Cellular Senescence (2021) · cites this paper as [54]

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Cited by

Sorted by citations

Authors
Benanti, J. A., & Galloway, D. A.
Affiliations
  • University of Washington (US)
  • Fred Hutch Cancer Center (US)
ORCID
Venue
Molecular and Cellular Biology
Publisher
Informa UK Limited
Published
2004-04-01
Volume
24
Issue
7
Pages
2842-2852
ISSN
  • 0270-7306 (ISSN-L)
  • 1098-5549
DOI
10.1128/mcb.24.7.2842-2852.2004
PMC
PMC371123
OpenAlex
W2081143567
Cited by
103
References
11
Citations by year
2026: 4 · 2025: 1 · 2024: 3 · 2023: 1 · 2022: 1 · 2021: 2 · 2020: 3 · 2019: 2 · 2018: 2 · 2017: 4 · 2016: 2 · 2015: 1 · 2014: 4 · 2013: 3 · 2012: 8
FWCI
7.61
Type
journal article
Language
en
Open access
Licence
journals.asm.org/non-commercial-tdm-license (tdm)
Locations
  • Molecular and Cellular Biology (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Cancer Institute (CA64795, R01CA064795)
  • National Institute of General Medical Sciences (GM-07270, T32-GM07270, T32GM007270)
Topics
  • Health Sciences › Medicine › Physiology › Telomeres, Telomerase, and Senescence
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Aging › Genetics, Aging, and Longevity in Model Organisms
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
Keywords
oncogenic Ras, cellular senescence, human fibroblasts, p16INK4a, retinoblastoma protein, hTERT, anchorage-independent growth, contact inhibition, growth arrest, senescence-associated β-galactosidase, cell transformation, p53
MeSH terms
Animals, Cells, Cultured, DNA-Binding Proteins, Fibroblasts, Humans, Retroviridae, Tumor Suppressor Protein p53, Retinoblastoma Protein, Cellular Senescence, ras Proteins, Telomerase, Cyclin-Dependent Kinase Inhibitor p16, Cell Size

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