Clinical and Translational Oncology

, Volume 18, Issue 4, pp 398–404 | Cite as

Methylation of the DCLK1 promoter region in circulating free DNA and its prognostic value in lung cancer patients

  • T. Powrózek
  • P. Krawczyk
  • M. Nicoś
  • B. Kuźnar-Kamińska
  • H. Batura-Gabryel
  • J. Milanowski
Research Article

Abstract

Introduction

The possibility of detection of suppressor genes methylation in circulating free DNA (cf-DNA) of cancer patients and the lack of methylation in healthy individuals makes this epigenetic alternation an ideal diagnostic marker of neoplastic processes. Moreover, hypermethylation in several genes promoter was described as a biomarker of lung cancer. Methylation in the gene encoding doublecortin-like kinase 1 (DCLK1) is observed in patients with colorectal cancer and cholangiocarcinoma. However, there are no studies concerning DCLK1 methylation in lung cancer patients. The aims of the study was to evaluate the frequency of DCLK1 promoter methylation in cf-DNA of lung cancer patients and of healthy persons as well as the usefulness of this test for predicting the lung cancer course.

Materials and methods

DCLK1 methylation status was evaluated in DNA isolated from peripheral blood plasma from 65 lung cancer patients and 95 healthy individuals. After DNA bisulfitation, DCLK1 methylation was determined using the qMSP-PCR technique. Moreover, the presence of DCLK1 methylation was correlated with the overall survival (OS) probability of lung cancer patients.

Results

DCLK1 promoter methylation was detected in 32 lung cancer patients (49.2 %) and 8 healthy individuals (8.4 %). The methylation of the region before transcription start site (TSS) and the region after TSS of DCLK1 gene was detected in 28 and 11 patients, respectively. In seven cases (10.8 %), the DCLK1 promoter methylation in both regions was reported simultaneously. The methylation was observed slightly frequently in patients with small cell lung cancer (75 % of SCLC patients). The median overall survival of patients with DCLK1 promoter methylation was lower than that of patients without DCLK1 gene modification (p = <0.001, HR = 4.235).

Conclusions

The evaluation of DCLK1 promoter region methylation may be useful in both early diagnosis and prediction of the course of lung cancer.

Keywords

Lung cancer DNA methylation DCLK1 Free-circulating DNA 

Notes

Acknowledgments

The study was sponsored by the Polpharma Scientific Foundation.

Compliance with ethical standards

Conflict of interest

None declared.

Informed consent

Informed consent was obtained from all individual participants included in the study. The research involved human participants.

References

  1. 1.
    Infante MV, Pedersen JH. Screening for lung cancer: are we there yet? Curr Opin Pulm Med. 2010;16:301–6.CrossRefPubMedGoogle Scholar
  2. 2.
    Boiselle PM. Computed tomography screening for lung cancer. JAMA. 2013;309:1163–70.CrossRefPubMedGoogle Scholar
  3. 3.
    Dela Cruz CS, Tanoue LT, Matthay RA. Lung cancer: epidemiology, etiology, and prevention. Clin Chest Med. 2011;32:605–44.CrossRefPubMedGoogle Scholar
  4. 4.
    Das PM, Singal R. DNA methylation and cancer. J Clin Oncol. 2004;22:4632–42.CrossRefPubMedGoogle Scholar
  5. 5.
    Ruth EB, Knight L, Greystoke A, Blackhall FH, Hughes A, Dive C, et al. DNA methylation in circulating tumour DNA as a biomarker for cancer. Biomark Insights. 2007;2:307–19.Google Scholar
  6. 6.
    Melissa A, Edwards, Pashayar P, Neavin DR, Brown MA. Methylation in tumorigenesis, methylation—from DNA, RNA and histones to diseases and treatment. Prof. Anica Dricu (Ed.), InTech 2012; 119–135. ISBN: 978-953-51-0881-8.Google Scholar
  7. 7.
    Jin H, Ma Y, Shen Q, Wang X. Circulating methylated DNA as biomarkers for cancer detection, methylation—from DNA, RNA and histones to diseases and treatment. Prof. Anica Dricu (Ed.), InTech 2012; 137–152, ISBN: 978-953-51-0881-8.Google Scholar
  8. 8.
    Nakanishi Y, Seno H, Fukuoka A, Ueo T, Yamaga Y, Maruno T, et al. Dclk1 distinguishes between tumor and normal stem cells in the intensine. Nat Genet. 2013;45:98–103.CrossRefPubMedGoogle Scholar
  9. 9.
    Vedeld HM, Skotheim RI, Lothe RA, Lind GE. The recently suggested intestinal cancer stem cell marker DCLK1 is an epigenetic biomarker for colorectal cancer. Epigenetics. 2014;9(3):346–50.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Andersen K, Boberg KM, Vedeld HM, Honne H, Hektoen M, Wadsworth CA, et al. Novel target genes and valid biomarker panel identified for cholangiocarcinoma. Epigenetics. 2012;7(11):1249–57.CrossRefGoogle Scholar
  11. 11.
    Dreos R, Ambrosini G, Périer R, Bucher P. EPD and EPDnew, high-quality promoter resources in the next-generation sequencing era. Nucl Acids Res. 2012;41:157–64.CrossRefGoogle Scholar
  12. 12.
    Gu C, Lu J, Cui T, Lu C, Shi H, Xu W, et al. Association between MGMT promoter methylation and non-small cell lung cancer: a meta-analysis. PLoS One. 2013;8(9):e72633.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Deep JS, Sidhu S, Chandel A, Thapliyal S, Garg C. Aberrant methylation in promoters of GSTP1, p16, p14, and RASSF1A Genes in smokers of North India. ISRN Pulmonol. 2012;2012: Art. ID 247631. doi:10.5402/2012/247631
  14. 14.
    Vaissière T, Hung RJ, Zaridze D, Moukeria A, Cuenin C, Fasolo V, et al. Quantitative analysis of DNA methylation profiles in lung cancer identifies aberrant DNA Methylation of specific genes and its association with gender and cancer risk factors. Cancer Res. 2009;69:243–52.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Lokk K, Vooder T, Kolde R, Valk K, Vosa U, Roosipuu R, et al. Methylation markers of early stage non-small cell lung cancer. PLoS One. 2012;7(6):e39813.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Wasserkort R, Kalmar A, Valcz G, Spisak S, Krispin M, Toth K, et al. Aberrant septin 9 DNA methylation in colorectal cancer is restricted to a single CpG island. BMC Cancer. 2013;30(13):398.CrossRefGoogle Scholar
  17. 17.
    Bañez LL, Sun L, Leenders GJ, Wheeler TM, Bangma CH, Freedland SJ, et al. Multicenter clinical validation of PITX2 Methylation as a prostate specific antigen Recurrence predictor in patients with post-radical prostatectomy prostate cancer. J Urol. 2010;184:149–56.CrossRefPubMedGoogle Scholar
  18. 18.
    Kim YJ, Yoon HY, Kim SK, Kim YW, Kim EJ, Kim YI, et al. EFEMP1 as a novel DNA methylation marker for prostate cancer: array-based DNA methylation and expression profiling. Clin Cancer Res. 2011;17(13):4523–30.CrossRefPubMedGoogle Scholar
  19. 19.
    Warren JD, Xiong W, Bunker AM, Vaughn CP, Furtado LV, Roberts WL, et al. Septin 9 methylated DNA is a sensitive and specific blood test for colorectal cancer. BMC Med. 2011;9:133.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Powrózek T, Krawczyk P, Kucharczyk T, Milanowski J. Septin 9 promoter region methylation in free circulating DNA-potential role in noninvasive diagnosis of lung cancer: preliminary report. Med Oncol. 2014;31(4):917.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Wu D, Xiong L, Wu S, Jiang M, Lian G, Wang M. TFPI-2 methylation predicts poor prognosis in non-small cell lung cancer. Lung Cancer. 2012;76:106–11.CrossRefPubMedGoogle Scholar
  22. 22.
    Niklinska W, Naumnik W, Sulewska A, Kozłowski M, Pankiewicz W, Milewski R. Prognostic significance of DAPK and RASSF1A promoter hypermethylation in non-small cell lung cancer (NSCLC). Folia Histochem Cytobiol. 2009;47(2):275–80.PubMedGoogle Scholar
  23. 23.
    Sandoval J, Mendez-Gonzalez J, Nadal E, Chen G, Carmona FJ, Sayols S, et al. A prognostic DNA methylation signature for stage I non-small-cell lung cancer. J Clin Oncol. 2013;31(32):4140–7.CrossRefPubMedGoogle Scholar

Copyright information

© Federación de Sociedades Españolas de Oncología (FESEO) 2015

Authors and Affiliations

  • T. Powrózek
    • 1
  • P. Krawczyk
    • 1
  • M. Nicoś
    • 1
  • B. Kuźnar-Kamińska
    • 2
  • H. Batura-Gabryel
    • 2
  • J. Milanowski
    • 1
  1. 1.Department of Pneumonology, Oncology and AllergologyMedical University of LublinLublinPoland
  2. 2.Department of Pulmonology, Allergology and Respiratory OncologyPoznań University of Medical SciencesPoznańPoland

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