DNA Hypermethylation af a Panel Of Genes as an Urinary Biomarker For Bladder Cancer Diagnosis
Purpose: Several studies have shown frequent changes in DNA methylation in bladder cancer (BCa), which vary among different geographical areas. The aim of this study is to examine the diagnostic accuracy of a panel of DNA methylation biomarkers in a Greek clinical setting contributing to the development of a universal panel of urine biomarkers.
Materials and Methods: Individuals with primary BCa and control individuals matching the gender, age and smoking status of the cancer patients were recruited. DNA methylation was assessed for the gene promoters of RASSF1, RARB, DAPK, TERT and APC in urine samples collected by spontaneous urination using quantitative Methylation Specific PCR (qMSP). All genes had been previously separately associated with BCa.
Results: Fifty patients and 35 healthy controls were recruited, with average age of 70.26 years and average smoking status of 44.78 pack-years. In the BCa group, DNA methylation was detected in 27 (61.4%) samples. RASSF1 was methylated in 52.2% of samples. Only 3 (13.6%) samples from the control group were methylated, all in the RASSF1 gene promoter. The specificity and sensitivity of this panel of genes to diagnose BCa was 86% and 61% respectively. The RASSF1 gene could diagnose BCa with specificity 86.4% and sensitivity 52.3%.
Conclusion: Promoter DNA methylation of this panel of five genes could be further investigated as urine biomarker for the diagnosis of BCa. The RASSF1 could be a single candidate biomarker for predicting BCa patients versus controls. Studies are required in order to develop a geographically adjusted diagnostic biomarker for BCa.
- Urinary Bladder Neoplasms; DNA Methylation; RASSF1; Urine Biomarker
How to Cite
2. Yafi FA, Brimo F, Steinberg J, Aprikian AG, Tanguay S, Kassouf W. Prospective analysis of sensitivity and specificity of urinary cytology and other urinary biomarkers for bladder cancer. Urol Oncol. 2015;33(2):66.e25-66.e6.6E31.
3. Barkan GA, Wojcik EM, Nayar R, et al. (2016) The Paris system for reporting urinary cytology: the quest to develop a standardized terminology. Acta Cytol 60:185–197
4. Chung W, Bondaruk J, Jelinek J, et al. Detection of bladder cancer using novel DNA methylation biomarkers in urine sediments. Cancer Epidemiol Biomarkers Prev. 2011;20(7):1483-91.
5. Das PM, Singal R. DNA methylation and cancer. J Clin Oncol. 2004;22(22):4632-42.
6. Li HT, Duymich CE, Weisenberger DJ, Liang G. Genetic and Epigenetic Alterations in Bladder Cancer. Int Neurourol J. 2016;20(Suppl 2):S84-94.
7. Pietrusiński M, Kȩpczyński Ƚ, Jȩdrzejczyk A, et al. Detection of bladder cancer in urine sediments by a hypermethylation panel of selected tumor suppressor genes. Cancer Biomark. 2017;18(1):47-59.
8. Phé V, Cussenot O, Rouprêt M. Interest of methylated genes as biomarkers in urothelial cell carcinomas of the urinary tract. BJU Int. 2009;104(7):896-901.
9. Leão R, Lee D, Figueiredo A, et al. Combined genetic and epigenetic alterations of the TERT promoter affect clinical and biological behavior of bladder cancer. Int J Cancer. 2019;144(7):1676-1684.
10. Zhan L, Zhang B, Tan Y, et al. Quantitative assessment of the relationship between RASSF1A gene promoter methylation and bladder cancer (PRISMA). Medicine (Baltimore). 2017;96(7):e6097.
11. Dai L, Ma C, Zhang Z, et al. DAPK Promoter Methylation and Bladder Cancer Risk: A Systematic Review and Meta-Analysis. PLoS One. 2016;11(12):e0167228.
12. Andrés G, Ashour N, Sánchez-Chapado M, Ropero S, Angulo JC. The study of DNA methylation in urological cancer: present and future. Actas Urol Esp. 2013;37(6):368‐375.
13. Kader F, Ghai M. DNA methylation-based variation between human populations. Mol Genet Genomics. 2017;292(1):5–35.
14. Friedrich MG, Weisenberger DJ, Cheng JC, et al. Detection of methylated apoptosis-associated genes in urine sediments of bladder cancer patients. Clin Cancer Res. 2004;10(22):7457-65.
15. Hoque MO, Feng Q, Toure P, et al. Detection of aberrant methylation of four genes in plasma DNA for the detection of breast cancer. J Clin Oncol. 2006; 24(26):4262-9.
16. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68(6):394-424.
17. Herr HW. Tumor progression and survival of patients with high grade, noninvasive papillary (TaG3) bladder tumors: 15-year outcome. J Urol. 2000; 163:60–2.
18. Beukers W, Kandimalla R, van Houwelingen D, et al.The use of molecular analyses in voided urine for the assessment of patients with hematuria. PLoS One. 2013;8(10):e77657
19. Renard I, Joniau S, van Cleynenbreugel B, et al. Identification and validation of the methylated TWIST1 and NID2 genes through real-time methylation-specific polymerase chain reaction assays for the noninvasive detection of primary bladder cancer in urine samples. Eur Urol. 2010 Jul;58(1):96-104.
20. Chen H, Yu Y, Rong S, Wang H. Evaluation of diagnostic accuracy of DNA methylation biomarkers for bladder cancer: a systematic review and meta-analysis. Biomarkers. 2014 May;19(3):189-97.
21. Zhang N, Chen S, Wu L, et al. Identification of Cancer-Specific Methylation of Gene Combination for the Diagnosis of Bladder Cancer. J Cancer. 2019;10(26):6761–6766.
22. van der Heijden AG, Mengual L, Ingelmo-Torres M, et al. Urine cell-based DNA methylation classifier for monitoring bladder cancer. Clin Epigenetics. 2018;10:71.
23. Giuliani C, Sazzini M, Bacalini MG, et al. Epigenetic Variability across Human Populations: A Focus on DNA Methylation Profiles of the KRTCAP3, MAD1L1 and BRSK2 Genes. Genome Biol Evol. 2016;8(9):2760-73.
24. Negraes PD, Favaro FP, Camargo JL, et al. DNA methylation patterns in bladder cancer and washing cell sediments: a perspective for tumor recurrence detection. BMC Cancer. 2008;8:238.
25. Pfeifer GP, Dammann R. Methylation of the tumor suppressor gene RASSF1A in human tumors. Biochemistry 2005;70:576–83.
26. Beukers W, Kandimalla R, Masius RG, et al. Stratification based on methylation of TBX2 and TBX3 into three molecular grades predicts progression in patients with pTa-bladder cancer. Mod Pathol. 2015;28(4):515-22.
27. Wolff EM, Chihara Y, Pan F, et al. Unique DNA methylation patterns distinguish noninvasive and invasive urothelial cancers and establish an epigenetic field defect in premalignant tissue. Cancer Res. 2010;70(20):8169-78.
28. Zuiverloon TC, Tjin SS, Busstra M, Bangma CH, Boevé ER, Zwarthoff EC. Optimization of nonmuscle invasive bladder cancer recurrence detection using a urine based FGFR3 mutation assay. J Urol. 2011;186(2):707‐712.
29. Larsen LK, Lind GE, Guldberg P, Dahl C. DNA-Methylation-Based Detection of Urological Cancer in Urine: Overview of Biomarkers and Considerations on Biomarker Design, Source of DNA, and Detection Technologies. Int J Mol Sci. 2019;20(11):2657.
30. Beltrán-García J, Osca-Verdegal R, Mena-Mollá S, García-Giménez JL. Epigenetic IVD Tests for Personalized Precision Medicine in Cancer. Front Genet. 2019;10:621.
- Abstract Viewed: 0 times
- Just Accepted/6743 Downloaded: 0 times