Breast Cancer Research and Treatment

, Volume 135, Issue 3, pp 705–713 | Cite as

Diagnostic values of GHSR DNA methylation pattern in breast cancer

  • Sandeep Kumar Botla
  • Amin Moghaddas Gholami
  • Mahdi Malekpour
  • Evgeny A. Moskalev
  • Mahdi Fallah
  • Pouria Jandaghi
  • Ali Aghajani
  • Irina S. Bondar
  • Ramesh Omranipour
  • Fatemeh Malekpour
  • Abbas Mohajeri
  • Azin Jahangiri Babadi
  • Özgür Sahin
  • Vladimir V. Bubnov
  • Hossein Najmabadi
  • Jörg D. Hoheisel
  • Yasser Riazalhosseini
Preclinical Study


DNA methylation patterns have been recognised as cancer-specific markers with high potential for clinical applications. We aimed at identifying methylation variations that differentiate between breast cancers and other breast tissue entities to establish a signature for diagnosis. Candidate genomic loci were analysed in 117 fresh-frozen breast specimens, which included cancer, benign and normal breast tissues from patients as well as material from healthy individuals. A cancer-specific DNA methylation signature was identified by microarray analysis in a test set of samples (n = 52, p < 2.1 × 10−4) and its performance was assessed through bisulphite pyrosequencing in an independent validation set (n = 65, p < 1.9 × 10−7). The signature is associated with SFRP2 and GHSR genes, and exhibited significant hypermethylation in cancers. Normal-appearing breast tissues from cancer patients were also methylated at these loci but to a markedly lower extent. This occurrence of methylated DNA in normal breast tissue of cancer patients is indicative of an epigenetic field defect. Concerning diagnosis, receiver operating characteristic curves and the corresponding area under the curve (AUC) analysis demonstrated a very high sensitivity and specificity of 89.3 and 100 %, respectively, for the GHSR methylation pattern (AUC >0.99). To date, this represents the DNA methylation marker of the highest sensitivity and specificity for breast cancer diagnosis. Functionally, ectopic expression of GHSR in a cell line model reduced breast cancer cell invasion without affecting cell viability upon stimulation of cells with ghrelin. Our data suggest a link between epigenetic down-regulation of GHSR and breast cancer cell invasion.


DNA methylation Breast cancer Diagnosis GHSR Epigenetics 



This study was supported by Bundesministerium für Bildung und Forschung as part of the NGFN programme (Grant Number 01GS08117). The authors thank Verena Beier and Neeme Tõnisson for discussions, and Achim Stephan for technical support.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10549_2012_2197_MOESM1_ESM.doc (1 mb)
Supplementary material 1 (DOC 1026 kb)


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Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Sandeep Kumar Botla
    • 1
  • Amin Moghaddas Gholami
    • 2
  • Mahdi Malekpour
    • 3
    • 4
  • Evgeny A. Moskalev
    • 1
  • Mahdi Fallah
    • 5
  • Pouria Jandaghi
    • 1
    • 6
  • Ali Aghajani
    • 7
  • Irina S. Bondar
    • 8
  • Ramesh Omranipour
    • 3
  • Fatemeh Malekpour
    • 9
  • Abbas Mohajeri
    • 10
  • Azin Jahangiri Babadi
    • 1
  • Özgür Sahin
    • 11
  • Vladimir V. Bubnov
    • 12
  • Hossein Najmabadi
    • 7
  • Jörg D. Hoheisel
    • 1
  • Yasser Riazalhosseini
    • 1
    • 13
  1. 1.Division of Functional Genome AnalysisGerman Cancer Research Center (DKFZ)HeidelbergGermany
  2. 2.Chair of Proteomics and BioanalyticsTechnische Universität MünchenFreisingGermany
  3. 3.Imam Khomeini Hospital, Cancer InstituteTehran University of Medical SciencesTehranIran
  4. 4.Department of MedicineVanderbilt University Medical CenterNashvilleUSA
  5. 5.Division of Molecular Genetic EpidemiologyGerman Cancer Research Center, (DKFZ)HeidelbergGermany
  6. 6.Research Center of Medical SciencesI.A.U, Tehran Medical BranchTehranIran
  7. 7.Genetics Research CenterUniversity of Social Welfare and Rehabilitation SciencesTehranIran
  8. 8.Regional Oncology ClinicOdessaUkraine
  9. 9.Shohada Tajrish HospitalShahid Beheshti University of Medical SciencesTehranIran
  10. 10.Department of Research and DevelopmentZahravi Pharmaceutical CompanyTabrizIran
  11. 11.Division of Molecular Genome AnalysisGerman Cancer Research Center (DKFZ)HeidelbergGermany
  12. 12.Department of Genomics and ImmunologyOdessa State Medical UniversityOdessaUkraine
  13. 13.Department of Human GeneticsMcGill University and Genome Quebec Innovation CentreMontrealCanada

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