Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Ha-RasG12V induces senescence in primary and immortalized human esophageal keratinocytes with p53 dysfunction

Abstract

Oncogenic Ras induces premature senescence in primary cells. Such an oncogene-induced senescence involves activation of tumor suppressor genes that provide a checkpoint mechanism against malignant transformation. In mouse, the ARF–p53 pathway mediates Ha-RasG12V-induced senescence, and p19ARF−/− and p53−/− cells undergo transformation upon Ras activation. In addition, mouse cells, unlike human cells, express constitutively active telomerase and have long telomeres. However, it is unclear how Ras activation affects human cells of epithelial origin with p53 mutation and/or telomerase activation. In order to address this question, Ha-RasG12V was expressed ectopically in primary as well as hTERT-immortalized human esophageal keratinocytes stably expressing dominant-negative p53 mutants. In human esophageal keratinocytes, we found that Ha-RasG12V induced senescence regardless of p53 status and telomerase activation. Ras activation resulted in changes of cellular morphology, activation of senescence-associated β-galactosidase, and suppression of cell proliferation, all coupled with reduction in the hyperphosphorylated form of the retinoblastoma protein (pRb). Furthermore, Ha-RasG12V upregulated p16INK4a and downregulated cyclin-dependent kinase Cdk4 in human esophageal keratinocytes. Thus, Ras-mediated senescence may involve distinct mechanisms between human and mouse cells. Inactivation of the pRb pathway may be necessary for Ras to overcome senescence and transform human esophageal epithelial cells.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 5
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  • Andl CD, Mizushima T, Nakagawa H, Oyama K, Harada H, Chruma K, Herlyn M and Rustgi AK . (2003). J. Biol. Chem., 278, 1824–1830.

  • Barbacid M . (1987). Annu. Rev. Biochem, 56, 779–827.

  • Bos JL . (1989). Cancer Res., 49, 4682–4689.

  • Brookes S, Rowe J, Ruas M, Llanos S, Clark PA, Lomax M, James MC, Vatcheva R., Bates S, Vousden KH, Parry D, Gruis N, Smit N, Bergman W and Peters G . (2002). EMBO J., 21, 2936–2945.

  • Concin N, Stimpfl M, Zeillinger C, Wolff U, Hefler L, Sedlak J, Leodolter S and Zeillinger R . (2003). Int. J. Oncol., 22, 51–57.

  • Delgado MD, Vaque JP, Arozarena I, Lopez-Ilasaca MA, Martinez C, Crespo P and Leon J . (2000). Oncogene, 19, 783–790.

  • Deng Q, Liao R, Wu BL and Sun P . (2004). J. Biol. Chem., 279, 1050–1059.

  • Dimri GP, Itahana K, Acosta M and Campisi J . (2000). Mol. Cell. Biol., 20, 273–285.

  • Drayton S, Rowe J, Jones R, Vatcheva R, Cuthbert-Heavens D, Marshall J, Fried M and Peters G . (2003). Cancer Cell, 4, 301–310.

  • Ferbeyre G, de Stanchina E, Lin AW, Querido E, McCurrach ME, Hannon GJ and Lowe SW . (2002). Mol. Cell. Biol., 22, 3497–3508.

  • Ferbeyre G, de Stanchina E, Querido E, Baptiste N, Prives C and Lowe SW . (2000). Genes Dev., 14, 2015–2027.

  • Fisher GH, Wellen SL, Klimstra D, Lenczowski JM, Tichelaar JW, Lizak MJ, Whitsett JA, Koretsky A and Varmus HE . (2001). Genes Dev., 15, 3249–3262.

  • Franza Jr BR, Maruyama K, Garrels JI and Ruley HE . (1986). Cell, 44, 409–418.

  • Gadducci A, Cosio S, Muraca S and Genazzani AR . (2002). Eur. J. Gynaecol. Oncol., 23, 390–396.

  • Guerra C, Mijimolle N, Dhawahir A, Dubus P, Barradas M, Serrano M, Campuzano V and Barbacid M . (2003). Cancer Cell, 4, 111–120.

  • Haapajarvi T, Kivinen L, Heiskanen A, des Bordes C, Datto MB, Wang XF and Laiho M . (1999). Exp. Cell Res., 248, 272–279.

  • Harada H, Nakagawa H, Oyama K, Takaoka M, Andl CD, Jacobmeier B, von Werder A, Enders GH, Opitz OG and Rustgi AK . (2003). Mol. Cancer Res., 1, 729–738.

  • Hingorani SR, Petricoin EF, Maitra A, Rajapakse V, King C, Jacobetz MA, Ross S, Conrads TP, Veenstra TD, Hitt BA, Kawaguchi Y, Johann D, Liotta LA, Crawford HC, Putt ME, Jacks T, Wright CV, Hruban RH, Lowy AM and Tuveson DA . (2003). Cancer Cell, 4, 437–450.

  • Hollstein MC, Smits AM, Galiana C, Yamasaki H, Bos JL, Mandard A, Partensky C and Montesano R . (1988). Cancer Res., 48, 5119–5123.

  • Huot TJ, Rowe J, Harland M, Drayton S, Brookes S, Gooptu C, Purkis P, Fried M, Bataille V, Hara E, Newton-Bishop J and Peters G . (2002). Mol. Cell. Biol., 22, 8135–8143.

  • Itakura Y, Sasano H, Shiga C, Furukawa Y, Shiga K, Mori S and Nagura H . (1994). Cancer, 74, 795–804.

  • Johnson M, Dimitrov D, Vojta PJ, Barrett JC, Noda A, Pereira-Smith OM and Smith JR . (1994). Mol. Carcinog., 11, 59–64.

  • Kamijo T, Zindy F, Roussel MF, Quelle DE, Downing JR, Ashmun RA, Grosveld G and Sherr CJ . (1997). Cell, 91, 649–659.

  • Krimpenfort P, Quon KC, Mooi WJ, Loonstra A and Berns A . (2001). Nature, 413, 83–86.

  • Lazarov M, Kubo Y, Cai T, Dajee M, Tarutani M, Lin Q, Fang M, Tao S, Green CL and Khavari PA . (2002). Nat. Med., 8, 1105–1114.

  • Lin AW, Barradas M, Stone JC, van Aelst L, Serrano M and Lowe SW . (1998). Genes Dev., 12, 3008–3019.

  • Lin AW and Lowe SW . (2001). Proc. Natl. Acad. Sci. USA, 98, 5025–5030.

  • Mallette FA, Goumard S, Gaumont-Leclerc MF, Moiseeva O and Ferbeyre G . (2004). Oncogene, 23, 91–99.

  • Morgenstern JP and Land H . (1990). Nucleic Acids Res., 18, 3587–3596.

  • Nakagawa H, Katzka D and Rustgi AK . (2003). Gastrointestinal Cancers Rustgi, AK (ed). Elsevier: London, pp. 241–251.

    Google Scholar 

  • Newbold RF and Overell RW . (1983). Nature, 304, 648–651.

  • Opitz OG, Harada H, Suliman Y, Rhoades B, Sharpless NE, Kent R, Kopelovich L, Nakagawa H and Rustgi AK . (2002). J. Clin. Invest., 110, 761–769.

  • Palmero I, Pantoja C and Serrano M . (1998). Nature, 395, 125–126.

  • Pearson M, Carbone R, Sebastiani C, Cioce M, Fagioli M, Saito S, Higashimoto Y, Appella E, Minucci S, Pandolfi PP and Pelicci PG . (2000). Nature, 406, 207–210.

  • Ruley HE . (1990). Cancer Cells, 2, 258–268.

  • Russo T, Zambrano N, Esposito F, Ammendola R, Cimino F, Fiscella M, Jackman J, O'Connor PM, Anderson CW and Appella E . (1995). J. Biol. Chem., 270, 29386–29391.

  • Schreiber FS, Deramaudt TB, Brunner TB, Boretti MI, Gooch KJ, Stoffers DA, Bernhard EJ and Rustgi AK . (2004). Gastroenterology, 127, 250–260.

  • Serrano M, Lin AW, McCurrach ME, Beach D and Lowe SW . (1997). Cell, 88, 593–602.

  • Sharpless NE, Bardeesy N, Lee KH, Carrasco D, Castrillon DH, Aguirre AJ, Wu EA, Horner JW and DePinho RA . (2001). Nature, 413, 86–91.

  • Sigal A and Rotter V . (2000). Cancer Res., 60, 6788–6793.

  • Suliman Y, Opitz OG, Avadhani A, Burns TC, El-Deiry W, Wong DT and Rustgi AK . (2001). Cancer Res., 61, 6467–6473.

  • Suzuki H, Kurita M, Mizumoto K, Nishimoto I, Ogata E and Matsuoka M . (2003). Biochem. Biophys. Res. Commun., 312, 1273–1277.

  • Tuveson D, Shaw A, Willis N, Silver D, Jackson E, Chang S, Mercer K,, Grochow R, Hock H, Crowley D, Hingorani S, Zaks T, King C, Jacobetz M, Wang L, Bronson R, Orkin S, DePinho R and Jacks T . (2004). Cancer Cell, 5, 375–387.

  • van Oijen MG and Slootweg PJ . (2000). Clin. Cancer Res., 6, 2138–2145.

  • Wei W, Hemmer RM and Sedivy JM . (2001). Mol. Cell. Biol., 21, 6748–6757.

  • Weinberg RA . (1989). Cancer Res., 49, 3713–3721.

  • Weinberg RA . (1997). Cell, 88, 573–575.

  • Weinberg WC, Azzoli CG, Kadiwar N and Yuspa SH . (1994). Cancer Res., 54, 5584–5592.

  • Yano H, Shiozaki H, Kobayashi K, Yano T, Tahara H, Tamura S and Mori T . (1991). Cancer, 67, 91–98.

  • Yu Q, Geng Y and Sicinski P . (2001). Nature, 411, 1017–1021.

  • Zhu J, Woods D, McMahon M and Bishop JM . (1998). Genes Dev., 12, 2997–3007.

  • Zou X, Ray D, Aziyu A, Christov K, Boiko AD, Gudkov AV and Kiyokawa H . (2002). Genes Dev., 16, 2923–2934.

Download references

Acknowledgements

We are grateful to Drs Meenhard Herlyn and Anil K Rustgi for their support and advice. We thank Dr Scott W Lowe for providing the pBabe-Ha-Ras plasmid. This work was in part supported by NIH Grant R21 DK64249 and AGA/FDHN Research Scholar Award (HN), NCI Grant P01-CA098101 (HN, CA, KO) and its core facilities, AACR basic science research award (CNJ), The National Pancreas Foundation (TD) and the NIH/NIDDK Center for Molecular Studies in Digestive and Liver Diseases and its Morphology Core, Molecular Biology and Cell Culture Core facilities, and pilot grant program (P30 DK50306).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Hiroshi Nakagawa.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Takaoka, M., Harada, H., Deramaudt, T. et al. Ha-RasG12V induces senescence in primary and immortalized human esophageal keratinocytes with p53 dysfunction. Oncogene 23, 6760–6768 (2004). https://doi.org/10.1038/sj.onc.1207923

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1207923

Keywords

This article is cited by

Search

Quick links