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.
Similar content being viewed by others
References
Infante MV, Pedersen JH. Screening for lung cancer: are we there yet? Curr Opin Pulm Med. 2010;16:301–6.
Boiselle PM. Computed tomography screening for lung cancer. JAMA. 2013;309:1163–70.
Dela Cruz CS, Tanoue LT, Matthay RA. Lung cancer: epidemiology, etiology, and prevention. Clin Chest Med. 2011;32:605–44.
Das PM, Singal R. DNA methylation and cancer. J Clin Oncol. 2004;22:4632–42.
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.
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.
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.
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.
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.
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.
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.
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.
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
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Acknowledgments
The study was sponsored by the Polpharma Scientific Foundation.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
None declared.
Informed consent
Informed consent was obtained from all individual participants included in the study. The research involved human participants.
Rights and permissions
About this article
Cite this article
Powrózek, T., Krawczyk, P., Nicoś, M. et al. Methylation of the DCLK1 promoter region in circulating free DNA and its prognostic value in lung cancer patients. Clin Transl Oncol 18, 398–404 (2016). https://doi.org/10.1007/s12094-015-1382-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12094-015-1382-z