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:

Proteomics analysis of H-RAS-mediated oncogenic transformation in a genetically defined human ovarian cancer model

Oncogene volume 24, pages 6174–6184 (2005)Cite this article

Abstract

RAS is a small GTP binding protein mutated in approximately 30% human cancer. Despite its important role in the initiation and progression of human cancer, the underlying mechanism of RAS-induced human epithelial transformation remains elusive. In this study, we probe the cellular and molecular mechanisms of RAS-mediated transformation, by profiling two human ovarian epithelial cell lines. One cell line was immortalized with SV40 T/t antigens and the human catalytic subunit of telomerase (T29), while the second cell line was transformed with an additional oncogenic rasV12 allele (T29H). In total, 32 proteins associated with RAS-mediated transformation have been identified using peptide mass fingerprinting. These protein targets are involved in several cellular pathways, including metabolism, redox balance, calcium signaling, apoptosis, and cellular methylation. One such target, the 40 kDa procaspase 4 is significantly upregulated at the protein level in RAS-transformed T29H cells, related directly to signaling through MEK, but not PI3 kinase. Cellular caspase 4 activity is, however, suppressed in the T29H cells, suggesting that the maturation process of caspase 4 is abrogated in RAS-transformed T29H cells. Consistent with this notion, transformed T29H cells were less susceptible to the toxic effects of anti-Fas antibody than were immortalized, nontransformed T29 cells, associated with less activation of caspase 4. This study demonstrates that functional proteomic analysis of a genetically defined cancer model provides a powerful approach toward systematically identifying cellular targets associated with oncogenic transformation.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

Abbreviations

2-DE:

two-dimensional electrophoresis

ACN:

acetonitrile

HOSE:

human ovarian surface epithelia cells

hTERT:

human telomerase catalytic subunit

MALDI-TOF:

matrix-assisted laser desorption/ionization time-of-flight

References

  • Allan LA, Morrice N, Brady S, Magee G, Pathak S and Clarke PR . (2003). Nat. Cell Biol., 5, 647–654.

  • Argiles JM and Azcon-Bieto J . (1988). Mol. Cell Biochem., 81, 3–17.

  • Balabanov S, Zimmermann U, Protzel C, Scharf C, Klebingat KJ and Walther R . (2001). Eur. J. Biochem., 268, 5977–5980.

  • Bao JJ, Le XF, Wang RY, Yuan J, Wang L, Atkinson EN, LaPushin R, Andreeff M, Fang B, Yu Y and Bast Jr RC . (2002). Cancer Res., 62, 7264–7272.

  • Berchuck A, Kamel A, Whitaker R, Kerns B, Olt G, Kinney R, Soper JT, Dodge R, Clarke-Pearson DL and Marks P . (1990). Cancer Res., 50, 4087–4091.

  • Bernard K, Litman E, Fitzpatrick JL, Shellman YG, Argast G, Polvinen K, Everett AD, Fukasawa K, Norris DA, Ahn NG and Resing KA . (2003). Cancer Res., 63, 6716–6725.

  • Blum H, Beier H and Gross HL . (1987). Electrophoresis, 8, 93–99.

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

  • Calaf G, Tahin Q, Alvarado ME, Estrada S, Cox T and Russo J . (1994). Breast Cancer Res. Treat., 29, 169–177.

  • Cao QJ, Jones JG and Li M . (2001). Int. J. Gynecol. Pathol., 20, 346–352.

  • Demoz M, Castino R, Dragonetti A, Raiteri E, Baccino FM and Isidoro C . (1999). J. Cell Biochem., 73, 370–378.

  • Elenbaas B, Spirio L, Koerner F, Fleming MD, Zimonjic DB, Donaher JL, Popescu NC, Hahn WC and Weinberg RA . (2001). Genes Dev., 15, 50–65.

  • Gemignani ML, Schlaerth AC, Bogomolniy F, Barakat RR, Lin O, Soslow R, Venkatraman E and Boyd J . (2003). Gynecol. Oncol., 90, 378–381.

  • Glading A, Chang P, Lauffenburger DA and Wells A . (2000). J. Biol. Chem., 275, 2390–2398.

  • Goll DE, Thompson VF, Li H, Wei W and Cong J . (2003). Physiol Rev., 83, 731–801.

  • Hahn WC, Counter CM, Lundberg AS, Beijersbergen RL, Brooks MW and Weinberg RA . (1999). Nature, 400, 464–468.

  • Hanahan D and Weinberg RA . (2000). Cell, 100, 57–70.

  • Jones MB, Krutzsch H, Shu H, Zhao Y, Liotta LA, Kohn EC and Petricoin III EF . (2002). Proteomics, 2, 76–84.

  • Kamada S, Washida M, Hasegawa J, Kusano H, Funahashi Y and Tsujimoto Y . (1997). Oncogene, 15, 285–290.

  • Kulkarni S, Saido TC, Suzuki K and Fox JE . (1999). J. Biol. Chem., 274, 21265–21275.

  • Land H, Parada LF and Weinberg RA . (1983). Nature, 304, 596–602.

  • Li G and Iyengar R . (2002). Proc. Natl. Acad. Sci. USA, 99, 13254–13259.

  • Lindahl R . (1992). Crit Rev. Biochem. Mol. Biol., 27, 283–335.

  • Liu J, Yang G, Thompson-Lanza JA, Glassman A, Hayes K, Patterson A, Marquez RT, Auersperg N, Yu Y, Hahn WC, Mills GB and Bast Jr RC . (2004). Cancer Res., 64, 1655–1663.

  • Losch A, Schindl M, Kohlberger P, Lahodny J, Breitenecker G, Horvat R and Birner P . (2004). Gynecol. Oncol., 92, 545–552.

  • Lundberg AS, Randell SH, Stewart SA, Elenbaas B, Hartwell KA, Brooks MW, Fleming MD, Olsen JC, Miller SW, Weinberg RA and Hahn WC . (2002). Oncogene, 21, 4577–4586.

  • Martin CA and Panja A . (2002). Am. J. Physiol Gastrointest. Liver Physiol., 282, G92–G104.

  • Matthaei KI and Williams JF . (1987). J. Natl. Cancer Inst., 79, 805–809.

  • Ostergaard M, Rasmussen HH, Nielsen HV, Vorum H, Orntoft TF, Wolf H and Celis JE . (1997). Cancer Res., 57, 4111–4117.

  • Park KS, Cho SY, Kim H and Paik YK . (2002). Int. J. Cancer, 97, 261–265.

  • Patton SE, Martin ML, Nelsen LL, Fang X, Mills GB, Bast RC, Jr and Ostrowski MC . (1998). Cancer Res., 58, 2253–2259.

  • Rich JN, Guo C, McLendon RE, Bigner DD, Wang XF and Counter CM . (2001). Cancer Res., 61, 3556–3560.

  • Rogers J, Khan M and Ellis J . (1990). Adv. Exp. Med. Biol., 269, 195–203.

  • Ruley HE . (1983). Nature, 304, 602–606.

  • Shevchenko A, Wilm M, Vorm O and Mann M . (1996). Anal. Chem., 68, 850–858.

  • Sieben NL, Macropoulos P, Roemen GM, Kolkman-Uljee SM, Jan FG, Houmadi R, Diss T, Warren B, Al Adnani M, De Goeij AP, Krausz T and Flanagan AM . (2004). J. Pathol., 202, 336–340.

  • Singer G, Oldt R, III, Cohen Y, Wang BG, Sidransky D, Kurman RJ and Shih I . (2003). J. Natl. Cancer Inst., 95, 484–486.

  • Varras MN, Sourvinos G, Diakomanolis E, Koumantakis E, Flouris GA, Lekka-Katsouli J, Michalas S and Spandidos DA . (1999). Oncology, 56, 89–96.

  • Watnick RS, Cheng YN, Rangarajan A, Ince TA and Weinberg RA . (2003). Cancer Cell, 3, 219–231.

  • Yang G, Thompson JA, Fang B and Liu J . (2003). Oncogene, 22, 5694–5701.

  • Young TW, Mei FC, Yang G, Thompson-Lanza JA, Liu J and Cheng X . (2004). Cancer Res., 64, 4577–4584.

  • Yu J, Boyapati A and Rundell K . (2001). Virology, 290, 192–198.

  • Yu LR, Zeng R, Shao XX, Wang N, Xu YH and Xia QC . (2000). Electrophoresis, 21, 3058–3068.

Download references

Acknowledgements

We are indebted to Dr Natalie Ahn (University of Colorado, Boulder) for her continuous support and insightful discussions. We thank Dr Tony Haag in the Biomedical Resource Facility (supported by NCI grant R24CA88317, NIEHS Center Grant ES06676 and N01-HV-28184) for mass spectrometry analysis. This work is supported by American Cancer Society Research Scholar Grant RSG-01-035-01-TBE and National Institute of Health Grant GM060170 to XC. JC is supported by an American Cancer Society Research Scholar Grant RSG-04-028-01-CCE.

Author information

Authors and Affiliations

  1. Department of Pharmacology and Toxicology, The University of Texas Medical Branch, 301 University Boulevard, Galveston, 77555-1031, TX, USA

    Travis Young, Fang Mei & Xiaodong Cheng

  2. Department of Pathology, The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, 77030, TX, USA

    Jinsong Liu

  3. Department of Experimental Therapeutics, The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, 77030, TX, USA

    Robert C Bast Jr

  4. Department of Human Biological Chemistry and Genetics, The University of Texas Medical Branch, 301 University Boulevard, Galveston, 77555-1031, TX, USA

    Alexander Kurosky

Authors
  1. Travis Young
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fang Mei
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jinsong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Robert C Bast Jr
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alexander Kurosky
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xiaodong Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xiaodong Cheng.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Young, T., Mei, F., Liu, J. et al. Proteomics analysis of H-RAS-mediated oncogenic transformation in a genetically defined human ovarian cancer model. Oncogene 24, 6174–6184 (2005). https://doi.org/10.1038/sj.onc.1208753

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1208753

Keywords

This article is cited by

Search

Advanced search

Quick links