Advances in Clinical and Experimental Medicine

Adv Clin Exp Med
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ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
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Advances in Clinical and Experimental Medicine

2019, vol. 28, nr 3, March, p. 355–360

doi: 10.17219/acem/84935

Publication type: original article

Language: English

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Creative Commons BY-NC-ND 3.0 Open Access

DNA methylation analysis of selected genes for the detection of early-stage lung cancer using circulating cell-free DNA

Zhiping Yang1,B,C,D,F, Weibo Qi2,B,C,F, Li Sun2,B,F, Hui Zhou2,B,D, Biliu Zhou2,B,D,F, Yi Hu3,A,C,E,F

1 Department of Oncology, The First Affiliated Hospital of Jiaxing University, Zhejiang, China

2 Department of Clinical Laboratory, The First Affiliated Hospital of Jiaxing University, Zhejiang, China

3 Department of Chest Surgery, The First Affiliated Hospital of Jiaxing University, Zhejiang, China

Abstract

Background. Lung cancer is still the deadliest cancer in the world, but early diagnosis cannot be achieved because of the limitations of diagnostic methods. DNA methylation has been proven to be a potentially powerful tool for cancer detection and diagnosis over the past decade.
Objectives. We explored whether free DNA methylation in plasma can be a reliable biomarker for noninvasive lung cancer detection.
Material and Methods. We detected the methylation of 8 genes in plasma-free DNA of patients with pulmonary space-occupying lesions using real-time quantitative methylation-specific polymerase chain reaction (QMSP). Among the 50 selected patients, 39 were confirmed using pathological analysis as having early lung cancer and 11 had an inflammatory pseudotumor.
Results. The QMSP detection showed that the methylation levels of 8 genes in the patients were significantly higher than in the non-lung cancer group. The methylation level of CALCA was the highest and the methylation level of HOXA9 was the lowest. Methylation of RASSF1A, CDKN2A and DLEC1 occured only in lung cancer patients, while methylation of CALCA, CDH13, PITX2, HOXA9, and WT1 occured not only in lung cancer patients, but also in non-lung cancers. The specificity reached 95~100%, whether for a single gene or overall, but the sensitivity was relatively low for each gene. The sensitivity can reach 72% if the methylation of any of the 8 genes is positive and the overall specificity was 91%. The positive and negative predictive values were 96% and 60%, respectively.
Conclusion. Quantitative detection of DNA methylation in plasma is a potential method for early diagnosis of lung cancer.

Key words

lung cancer, early diagnosis, QSMP, plasma-free DNA

References (28)

  1. Carter D. New global survey shows an increasing cancer burden. Am J Nurs. 2014;114(3):17.
  2. Nardi-Agmon I, Peled N. Exhaled breath analysis for the early detection of lung cancer: Recent developments and future prospects. Lung Cancer (Auckl). 2017;8:31–38.
  3. Fujikawa A, Takiguchi Y, Mizuno S, et al. Lung cancer screening: Comparison of computed tomography and X-ray. Lung Cancer. 2008;61(2):195–201.
  4. Das PM, Singal R. DNA methylation and cancer. J Clin Oncol. 2004;22(22):4632–4642.
  5. Agostini M, Pucciarelli S, Enzo MV, et al. Circulating cell-free DNA: A promising marker of pathologic tumor response in rectal cancer patients receiving preoperative chemoradiotherapy. Ann Surg Oncol. 2011;18(9):2461–2468.
  6. Anker P, Lyautey J, Lederrey C, Stroun M. Circulating nucleic acids in plasma or serum. Clin Chim Acta. 2001;313(1–2):143–146.
  7. De Mattos-Arruda L, Olmos D, Tabernero J. Prognostic and predictive roles for circulating biomarkers in gastrointestinal cancer. Future Oncol. 2011;7(12):1385–1397.
  8. Lou-Qian Z, Rong Y, Ming L, Xin Y, Feng J, Lin X. The prognostic value of epigenetic silencing of p16 gene in NSCLC patients: A systematic review and meta-analysis. PLoS One. 2013;8(1):e54970.
  9. Hwang JA, Lee BB, Kim Y, et al. HOXA9 inhibits migration of lung cancer cells and its hypermethylation is associated with recurrence in non-small cell lung cancer. Mol Carcinog. 2015;54(Suppl 1):E72–80.
  10. Dietrich D, Hasinger O, Liebenberg V, Field JK, Kristiansen G, Soltermann A. DNA methylation of the homeobox genes PITX2 and SHOX2 predicts outcome in non-small-cell lung cancer patients. Diagn Mol Pathol. 2012;21(2):93–104.
  11. Wrangle J, Machida EO, Danilova L, et al. Functional identification of cancer-specific methylation of CDO1, HOXA9 and TAC1 for the diagnosis of lung cancer. Clin Cancer Res. 2014;20(7):1856–1864.
  12. Nawaz I, Qiu X, Wu H, et al. Development of a multiplex methylation specific PCR suitable for (early) detection of non-small cell lung cancer. Epigenetics. 2014;9(8):1138–1148.
  13. Harden SV, Tokumaru Y, Westra WH, et al. Gene promoter hypermethylation in tumors and lymph nodes of stage I lung cancer patients. Clin Cancer Res. 2003;9(4):1370–1375.
  14. Müller HM, Widschwendter M. Methylated DNA as a possible screening marker for neoplastic disease in several body fluids. Expert Rev Mol Diagn. 2003;3(4):443–458.
  15. Esteller M, Sanchez-Cespedes M, Rosell R, Sidransky D, Baylin SB, Herman JG. Detection of aberrant promoter hypermethylation of tumor suppressor genes in serum DNA from non-small cell lung cancer patients. Cancer Res. 1999;59(1):67–70.
  16. Hoque MO, Begum S, Topaloglu O, et al. Quantitative detection of promoter hypermethylation of multiple genes in the tumor, urine and serum DNA of patients with renal cancer. Cancer Res. 2004;64(15):5511–5517.
  17. Ibanez de Caceres I, Battagli C, Esteller M, et al. Tumor cell-specific BRCA1 and RASSF1A hypermethylation in serum, plasma and peritoneal fluid from ovarian cancer patients. Cancer Res. 2004;64(18):6476–6481.
  18. Andreeva AV, Kutuzov MA. Cadherin 13 in cancer. Genes Chromosomes Cancer. 2010;49(9):775–790.
  19. Wu LS, Qian JY, Wang M, Yang H. Identifying the role of Wilms tumor 1 associated protein in cancer prediction using integrative genomic analyses. Mol Med Rep. 2016;14(3):2823–2831.
  20. Gu L, Zhao T, Lu XL, Qin GY. Identification of epigenetic aberrant promoter methylation of p16INK4A in serum for non-small cell lung cancer early diagnosis. J Med Res. 2013;42:163–166.
  21. Zhao Y, Zhou H, Ma K, et al. Abnormal methylation of seven genes and their associations with clinical characteristics in early stage non-small cell lung cancer. Oncol Lett. 2013;5(4):1211–1218.
  22. Morán A, Fernández-Marcelo T, Carro J, et al. Methylation profiling in non-small cell lung cancer: Clinical implications. Int J Oncol. 2012;40(3):739–746.
  23. Zhai X, Li SJ. Methylation of RASSF1A and CDH13 genes in individualized chemotherapy for patients with non-small cell lung cancer. Asian Pac J Cancer Prev. 2014;15(12):4925–4928.
  24. Pastuszak-Lewandoska D, Kordiak J, Antczak A, et al. Expression level and methylation status of three tumor suppressor genes, DLEC1, ITGA9 and MLH1, in non-small cell lung cancer. Med Oncol. 2016;33(7):75.
  25. Zhang Y, Wang R, Song H, et al. Methylation of multiple genes as a candidate biomarker in non-small cell lung cancer. Cancer Lett. 2011;303(1):21–28.
  26. Fischer JR, Ohnmacht U, Rieger N, et al. Prognostic significance of RASSF1A promoter methylation on survival of non-small cell lung cancer patients treated with gemcitabine. Lung Cancer. 2007;56(1):115–123.
  27. Wang J, Wang B, Chen X, Bi J. The prognostic value of RASSF1A promoter hypermethylation in non-small cell lung carcinoma: A systematic review and meta-analysis. Carcinogenesis. 2011;32(3):411–416.
  28. Li W, Deng J, Tang JX. Combined effects methylation of FHIT, RASSF1A and RARbeta genes on non-small cell lung cancer in the Chinese population. Asian Pac J Cancer Prev. 2014;15(13):5233–5237.