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.

  • Oncogenomics
  • Published:

Epigenetic regulation of the ras effector/tumour suppressor RASSF2 in breast and lung cancer

Oncogene volume 27, pages 1805–1811 (2008)Cite this article

Abstract

RASSF2 is a recently identified member of a class of novel tumour suppressor genes, all containing a ras-association domain. RASSF2 resides at 20p13, a region frequently lost in human cancers. In this report we investigated methylation status of the RASSF2 promoter CpG island in a series of breast, ovarian and non-small cell lung cancers (NSCLC). RASSF2 was frequently methylated in breast tumour cell lines (65%, 13/20) and in primary breast tumours (38%, 15/40). RASSF2 expression could be switched back on in methylated breast tumour cell lines after treatment with 5′-aza-2′deoxycytidine. RASSF2 was also frequently methylated in NSCLC tumours (44%, (22/50). The small number of corresponding normal breast and lung tissue DNA samples analysed were unmethylated. We also did not detect RASSF2 methylation in ovarian tumours (0/17). Furthermore no mutations were found in the coding region of RASSF2 in these ovarian tumours. We identified a highly conserved putative bipartite nuclear localization signal (NLS) and demonstrated that endogenous RASSF2 localized to the nucleus. Mutation of the putative NLS abolished the nuclear localization. RASSF2 suppressed breast tumour cell growth in vitro and in vivo, while the ability of NLS-mutant RASSF2 to suppress growth was much diminished. Hence we demonstrate that RASSF2 has a functional NLS that is important for its tumour suppressor gene function. Our data from this and a previous report indicate that RASSF2 is frequently methylated in colorectal, breast and NSCLC tumours. We have identified RASSF2 as a novel methylation marker for multiple malignancies and it has the potential to be developed into a valuable marker for screening several cancers in parallel using promoter hypermethylation profiles.

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

Similar content being viewed by others

References

  • Agathanggelou A, Cooper WN, Latif F . (2005). Role of the ras-association domain family 1 tumor suppressor gene in human cancers. Cancer Res 65: 3497–3508.

    Article  CAS  PubMed  Google Scholar 

  • Agathanggelou A, Honorio S, Macartney DP, Martinez A, Dallol A, Radar J et al. (2001). Methylation associated inactivation of RASSF1A from region 3p21.3 in lung, breast and ovarian tumours. Oncogene 20: 1509–1518.

    Article  CAS  PubMed  Google Scholar 

  • Akino K, Toyota M, Suzuki H, Mita H, Sasaki Y, Ohe-Toyota M et al. (2005). The Ras effector RASSF2 is a novel tumor-suppressor gene in human colorectal cancer. Gastroenterology 129: 156–169.

    Article  CAS  PubMed  Google Scholar 

  • Allen NP, Donninger H, Vos MD, Eckfeld K, Hesson L, Gordon L et al. (2007). RASSF6 is a novel member of the RASSF family of tumor suppressors. Oncogene; e-pub ahead of print: 2 April 2007.

  • Burbee DG, Forgacs E, Zochbauer-Muller S, Shivakumar L, Fong K, Gao BN et al. (2001). Epigenetic inactivation of RASSF1A in lung and breast cancers and malignant phenotype suppression. J Natl Cancer Inst 93: 691–699.

    Article  CAS  PubMed  Google Scholar 

  • Cox AD, Der CJ . (2003). The dark side of Ras: regulation of apoptosis. Oncogene 22: 8999–9006.

    Article  CAS  PubMed  Google Scholar 

  • Dallol A, Agathanggelou A, Tommasi S, Pfeifer GP, Maher ER, Latif F . (2005). Involvement of the RASSF1A tumor suppressor gene in controlling cell migration. Cancer Res 65: 7653–7659.

    Article  CAS  PubMed  Google Scholar 

  • Dammann R, Li C, Yoon JH, Chin PL, Bates S, Pfeifer GP . (2000). Epigenetic inactivation of a RAS association domain family protein from the lung tumour suppressor locus 3p21.3. Nat Genet 25: 315–319.

    Article  CAS  PubMed  Google Scholar 

  • Eckfeld K, Hesson L, Vos MD, Bieche I, Latif F, Clark GJ . (2004). RASSF4/AD037 is a potential ras effector/tumor suppressor of the RASSF family. Cancer Res 64: 8688–8693.

    Article  CAS  PubMed  Google Scholar 

  • Guo C, Tommasi S, Liu L, Yee JK, Dammann R, Pfeifer GP . (2007). RASSF1A is part of a complex similar to the Drosophila Hippo/Salvador/Lats tumor-suppressor network. Curr Biol 17: 700–705.

    Article  CAS  PubMed  Google Scholar 

  • Hesson L, Bieche I, Krex D, Criniere E, Hoang-Xuan K, Maher ER et al. (2004). Frequent epigenetic inactivation of RASSF1A and BLU genes located within the critical 3p21.3 region in gliomas. Oncogene 23: 2408–2419.

    Article  CAS  PubMed  Google Scholar 

  • Hesson LB, Cooper WN, Latif F . (2007). Evaluation of the 3p21.3 tumour-suppressor gene cluster. Oncogene; e-pub ahead of print: 28 May 2007.

  • Hesson L, Dallol A, Minna JD, Maher ER, Latif F . (2003). NORE1A, a homologue of RASSF1A tumour suppressor gene is inactivated in human cancers. Oncogene 22: 947–954.

    Article  CAS  PubMed  Google Scholar 

  • Hesson LB, Wilson R, Morton D, Adams C, Walker M, Maher ER et al. (2005). CpG island promoter hypermethylation of a novel Ras-effector gene RASSF2A is an early event in colon carcinogenesis and correlates inversely with K-ras mutations. Oncogene 24: 3987–3994.

    Article  CAS  PubMed  Google Scholar 

  • Irimia M, Fraga MF, Sanchez-Cespedes M, Esteller M . (2004). CpG island promoter hypermethylation of the Ras-effector gene NORE1A occurs in the context of a wild-type K-ras in lung cancer. Oncogene 23: 8695–8699.

    Article  CAS  PubMed  Google Scholar 

  • Kashuba VI, Li JF, Wang FL, Senchenko VN, Protopopov A, Malyukova A et al. (2004). RBSP3 (HYA22) is a tumor suppressor gene implicated in major epithelial malignancies. Proc Natl Acad Sci USA 101: 4906–4911.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lambros MB, Fiegler H, Jones A, Gorman P, Roylance RR, Carter NP et al. (2005). Analysis of ovarian cancer cell lines using array-based comparative genomic hybridization. J Pathol 205: 29–40.

    Article  CAS  PubMed  Google Scholar 

  • Lerman MI, Minna JD . (2000). The 630-kb lung cancer homozygous deletion region on human chromosome 3p21.3: identification and evaluation of the resident candidate tumor suppressor genes. Cancer Res 60: 6116–6133.

    CAS  PubMed  Google Scholar 

  • Malumbres M, Barbacid M . (2003). RAS oncogenes: the first 30 years. Nat Rev Cancer 3: 459–465.

    Article  CAS  PubMed  Google Scholar 

  • Park HW, Kang HC, Kim IJ, Jang SG, Kim K, Yoon HJ et al. (2007). Correlation between hypermethylation of the RASSF2A promoter and K-ras/BRAF mutations in microsatellite-stable colorectal cancers. Int J Cancer 120: 7–12.

    Article  CAS  PubMed  Google Scholar 

  • Protopopov AI, Li J, Winberg G, Gizatullin RZ, Kashuba VI, Klein G et al. (2002). Human cell lines engineered for tetracycline-regulated expression of tumor suppressor candidate genes from a frequently affected chromosomal region, 3p21. J Gene Med 4: 397–406.

    Article  CAS  PubMed  Google Scholar 

  • Robbins J, Dilworth SM, Laskey RA, Dingwall C . (1991). Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence. Cell 64: 615–623.

    Article  CAS  PubMed  Google Scholar 

  • Rommel C, Hafen E . (1998). Ras—a versatile cellular switch. Curr Opin Genet Dev 8: 412–418.

    Article  CAS  PubMed  Google Scholar 

  • Schubbert S, Bollag G, Shannon K . (2007). Deregulated Ras signaling in developmental disorders: new tricks for an old dog. Curr Opin Genet Dev 17: 15–22.

    Article  CAS  PubMed  Google Scholar 

  • Shields JM, Pruitt K, McFall A, Shaub A, Der CJ . (2000). Understanding Ras: ‘it ain't over ‘til it's over’. Trends Cell Biol 10: 147–154.

    Article  CAS  PubMed  Google Scholar 

  • Tommasi S, Dammann R, Jin SG, Zhang XF, Avruch J, Pfeifer GP . (2002). RASSF3 and NORE1: identification and cloning of two human homologues of the putative tumor suppressor gene RASSF1. Oncogene 21: 2713–2720.

    Article  CAS  PubMed  Google Scholar 

  • Tommasi S, Dammann R, Zhang ZQ, Wang Y, Liu LM, Tsark WM et al. (2005). Tumor susceptibility of Rassf1a knockout mice. Cancer Res 65: 92–98.

    CAS  PubMed  Google Scholar 

  • van der Weyden L, Tachibana KK, Gonzalez MA, Adams DJ, Ng BL, Petty R et al. (2005). The RASSF1A isoform of RASSF1 promotes microtubule stability and suppresses tumorigenesis. Mol Cell Biol 25: 8356–8367.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vos MD, Ellis CA, Elam C, Ulku AS, Taylor BJ, Clark GJ . (2003). RASSF2 is a novel K-Ras-specific effector and potential tumor suppressor. J Biol Chem 278: 28045–28051.

    Article  CAS  PubMed  Google Scholar 

  • Yoon JH, Damman R, Pfeifer GP . (2001). Hypermethylation of the CpG island of the RASSF1A gene in ovarian and renal cell carcinomas. Int J Cancer 94: 212–217.

    Article  CAS  PubMed  Google Scholar 

  • Zhang Z, Sun D, Van do N, Tang A, Hu L, Huang G . (2007). Inactivation of RASSF2A by promoter methylation correlates with lymph node metastasis in nasopharyngeal carcinoma. Int J Cancer 120: 32–38.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

Research in FL laboratory funded in part by Breast Cancer Campaign, Cancer Research UK, Association for International Cancer Research and Birmingham Children's Hospital Research Foundation. ERZ was supported by research grants from the Swedish Cancer Society, Swedish Research Council, Swedish Foundation for International Cooperation in Research and Higher Education (STINT), Swedish Institute, Royal Swedish Academy of Sciences, INTAS and Karolinska Institute.

Author information

Authors and Affiliations

  1. Department of Medical and Molecular Genetics, Division of Reproductive and Child Health, Institute of Biomedical Research, University of Birmingham, Edgbaston, Birmingham, UK

    W N Cooper, R E Dickinson, A Dallol, L B Hesson, E R Maher & F Latif

  2. MTC, Karolinska Institute, Theorells vag, Stockholm, Sweden

    E V Grigorieva, T V Pavlova & E R Zabarovsky

  3. Institute of Molecular Biology and Biophysics SD RAMS. Timakova 2, Novosibirsk, Russia

    E V Grigorieva

  4. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia

    T V Pavlova & E R Zabarovsky

  5. Laboratoire d'Oncogenetique-INSERM E0017, Centre Rene Huguenin, St-Cloud, France

    I Bieche

  6. Laboratory of Molecular Pharmacology, Istituto di Ricerche Farmacologiche ‘Mario Negri’, Milan, Italy

    M Broggini

  7. Department Of Medicine, Molecular Targets Group, J. G. Brown Cancer Center, University of Louisville, Louisville, KY, USA

    G J Clark

Authors
  1. W N Cooper
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R E Dickinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A Dallol
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. E V Grigorieva
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. T V Pavlova
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. L B Hesson
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. I Bieche
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. M Broggini
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. E R Maher
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. E R Zabarovsky
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. G J Clark
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. F Latif
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to F Latif.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cooper, W., Dickinson, R., Dallol, A. et al. Epigenetic regulation of the ras effector/tumour suppressor RASSF2 in breast and lung cancer. Oncogene 27, 1805–1811 (2008). https://doi.org/10.1038/sj.onc.1210805

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1210805

Keywords

This article is cited by

Search

Advanced search

Quick links