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Open Access 01-12-2022 | Biomarkers | Research

CALN1 hypomethylation as a biomarker for high-risk bladder cancer

Authors: Kimiaki Takagi, Azumi Naruse, Kazutoshi Akita, Yuka Muramatsu-Maekawa, Kota Kawase, Takuya Koie, Masanobu Horie, Arizumi Kikuchi

Published in: BMC Urology | Issue 1/2022

Abstract

Background

DNA methylation in cancer is considered a diagnostic and predictive biomarker. We investigated the usefulness of the methylation status of CALN1 as a biomarker for bladder cancer using methylation-sensitive restriction enzyme (MSRE)-quantitative polymerase chain reaction (qPCR).

Methods

Eighty-two bladder cancer fresh samples were collected via transurethral resection of bladder tumors. Genomic DNA was extracted from the samples, and MSRE-qPCR was performed to determine the CALN1 methylation percentage. Reverse transcription-qPCR was performed to assess the correlation between CALN1 methylation and mRNA expression. The association between CALN1 methylation percentage and clinicopathological variables of all cases and intravesical recurrence of non-muscle-invasive bladder cancer (non-MIBC) cases were analyzed.

Results

Of the 82 patients, nine had MIBC and 71 had non-MIBC who had not undergone total cystectomy. The median CALN1 methylation percentage was 79.5% (interquartile range: 51.1–92.6%). The CALN1 methylation percentage had a negative relationship with CALN1 mRNA expression (Spearman’s ρ = − 0.563 and P = 0.012). Hypomethylation of CALN1 was associated with advanced tumor stage (P = 0.0007) and histologically high grade (P = 0.018). Furthermore, multivariate analysis revealed that CALN1 hypomethylation was an independent risk factor for intravesical recurrence in non-MIBC patients (hazard ratio 3.83, 95% confidence interval; 1.14–13.0, P = 0.031).

Conclusion

Our findings suggest that CALN1 methylation percentage could be a useful molecular biomarker for bladder cancer.

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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;68:394–424. https://doi.org/10.3322/caac.21492.CrossRefPubMed

Cancer. Net. Bladder cancer: statistics. Cancer.Net, Doctor-approved patient information from ASCO website. 2022. https://www.cancer.net/cancer-types/bladder-cancer/statistics. Accessed 15 Aug 2022.

van Rhijn BW, Burger M, Lotan Y, Solsona E, Stief CG, Sylvester RJ, et al. Recurrence and progression of disease in non-muscle-invasive bladder cancer: from epidemiology to treatment strategy. Eur Urol. 2009;56:430–42. https://doi.org/10.1016/j.eururo.2009.06.028.CrossRefPubMed

Pajor G, Somogyi L, Melegh B, Alpar D, Kajtar B, Farkas L, et al. Urovysion: considerations on modifying current evaluation scheme, including immunophenotypic targeting and locally set, statistically derived diagnostic criteria. Cytometry A. 2011;79:375–82. https://doi.org/10.1002/cyto.a.21065.CrossRefPubMed

Raitanen MP, Marttila T, Nurmi M, Ala-Opas M, Nieminen P, Aine R, et al. Human complement factor H related protein test for monitoring bladder cancer. J Urol. 2001;165:374–7. https://doi.org/10.1097/00005392-200102000-00005.CrossRefPubMed

Landman J, Chang Y, Kavaler E, Droller MJ, Liu BC. Sensitivity and specificity of NMP-22, telomerase, and BTA in the detection of human bladder cancer. Urology. 1998;52:398–402. https://doi.org/10.1016/s0090-4295(98)00219-2.CrossRefPubMed

Pajor G, Sule N, Alpar D, Kajtar B, Kneif M, Bollmann D, et al. Increased efficiency of detecting genetically aberrant cells by UroVysion test on voided urine specimens using automated immunophenotypical preselection of uroepithelial cells. Cytom A. 2008;73:259–65. https://doi.org/10.1002/cyto.a.20528.CrossRef

Robertson AG, Kim J, Al-Ahmadie H, Bellmunt J, Guo G, Cherniack AD, et al. Comprehensive molecular characterization of muscle-invasive bladder cancer. Cell. 2017;171:540-56.e25. https://doi.org/10.1016/j.cell.2017.09.007.

Hegi ME, Diserens AC, Gorlia T, Hamou MF, de Tribolet N, Weller M, et al. MGMT gene silencing and benefit from temozolomide in glioblastoma. N Engl J Med. 2005;352:997–1003. https://doi.org/10.1056/NEJMoa043331.CrossRefPubMed

10.

deVos T, Tetzner R, Model F, Weiss G, Schuster M, Distler J, et al. Circulating methylated SEPT9 DNA in plasma is a biomarker for colorectal cancer. Clin Chem. 2009;55:1337–46. doi:https://doi.org/10.1373/clinchem.2008.115808.CrossRefPubMed

11.

Queirós AC, Villamor N, Clot G, Martinez-Trillos A, Kulis M, Navarro A, et al. A B-cell epigenetic signature defines three biologic subgroups of chronic lymphocytic leukemia with clinical impact. Leukemia. 2015;29:598–605. https://doi.org/10.1038/leu.2014.252.CrossRefPubMed

12.

Wu YQ, Lin X, Liu CM, Jamrich M, Shaffer LG. Identification of a human brain-specific gene, calneuron 1, a new member of the calmodulin superfamily. Mol Genet Metab. 2001;72:343–50. https://doi.org/10.1006/mgme.2001.3160.CrossRefPubMed

13.

Roderick HL, Cook SJ. Ca2 + signalling checkpoints in cancer: remodelling Ca2 + for cancer cell proliferation and survival. Nat Rev Cancer. 2008;8:361–75. https://doi.org/10.1038/nrc2374.CrossRefPubMed

14.

Shapovalov G, Ritaine A, Skryma R, Prevarskaya N. Role of TRP ion channels in cancer and tumorigenesis. Semin Immunopathol. 2016;38:357–69. https://doi.org/10.1007/s00281-015-0525-1.CrossRefPubMed

15.

Laird PW. The power and the promise of DNA methylation markers. Nat Rev Cancer. 2003;3:253–66. https://doi.org/10.1038/nrc1045.CrossRefPubMed

16.

Bock C. Epigenetic biomarker development. Epigenomics. 2009;1:99–110. https://doi.org/10.2217/epi.09.6.CrossRefPubMed

17.

Ohmura H, Ito M, Uchino K, Okada C, Tanishima S, Yamada Y, et al. Methylation of drug resistance-related genes in chemotherapy-sensitive Epstein–Barr virus-associated gastric cancer. FEBS Open Bio. 2020;10:147–57. https://doi.org/10.1002/2211-5463.12765.CrossRefPubMed

18.

Yu Y, Cao H, Zhang M, Shi F, Wang R, Liu X. Prognostic value of DNA methylation for bladder cancer. Clin Chim Acta. 2018;484:207–12. https://doi.org/10.1016/j.cca.2018.05.056.CrossRefPubMed

19.

Cao R, Meng Z, Liu T, Wang G, Qian G, Cao T, et al. Decreased TRPM7 inhibits activities and induces apoptosis of bladder cancer cells via ERK1/2 pathway. Oncotarget. 2016;7:72941–60. https://doi.org/10.18632/oncotarget.12146.CrossRefPubMedPubMedCentral

20.

Wang YM, Zhou P, Wang LY, Li ZH, Zhang YN, Zhang YX. Correlation between DNase I hypersensitive site distribution and gene expression in HeLa S3 cells. PLoS One. 2012;7:e42414. https://doi.org/10.1371/journal.pone.0042414.CrossRefPubMedPubMedCentral

21.

Guo RQ, Xiong GY, Yang KW, Zhang L, He SM, Gong YQ, et al. Detection of urothelial carcinoma, upper tract urothelial carcinoma, bladder carcinoma, and urothelial carcinoma with gross hematuria using selected urine-DNA methylation biomarkers: a prospective, single-center study. Urol Oncol. 2018;36:342.e15-23. https://doi.org/10.1016/j.urolonc.2018.04.001.CrossRef

22.

Reinert T, Modin C, Castano FM, Lamy P, Wojdacz TK, Hansen LL, et al. Comprehensive genome methylation analysis in bladder cancer: identification and validation of novel methylated genes and application of these as urinary tumor markers. Clin Cancer Res. 2011;17:5582–92. https://doi.org/10.1158/1078-0432.CCR-10-2659.CrossRefPubMed

23.

van der Heijden AG, Mengual L, Ingelmo-Torres M, Lozano JJ, vande Rijt-vanWesterlo CCM, Baixauli M, et al. Urine cell-based DNA methylation classifier for monitoring bladder cancer. Clin Epigenet. 2018;10:71. https://doi.org/10.1186/s13148-018-0496-x.CrossRef

24.

Millar D, Christova Y, Holliger P. A polymerase engineered for bisulfite sequencing. Nucleic Acids Res. 2015;43:e155. https://doi.org/10.1093/nar/gkv798.CrossRefPubMedPubMedCentral

25.

Perry N, Wasko K, Cheng J, Tabbaa D, Marco E, Giannoukos G, et al. Methylation-sensitive restriction enzyme quantitative polymerase chain reaction enables rapid, accurate, and precise detection of methylation status of the regulatory T cell (Treg)-specific demethylation region in primary human Tregs. J Immunol. 2021;206:446–51. https://doi.org/10.4049/jimmunol.1901275.CrossRefPubMed

Title: CALN1 hypomethylation as a biomarker for high-risk bladder cancer
Authors: Kimiaki Takagi
Azumi Naruse
Kazutoshi Akita
Yuka Muramatsu-Maekawa
Kota Kawase
Takuya Koie
Masanobu Horie
Arizumi Kikuchi
Publication date: 01-12-2022
Publisher: BioMed Central
Keyword: Biomarkers
Published in: BMC Urology / Issue 1/2022
Electronic ISSN: 1471-2490
DOI: https://doi.org/10.1186/s12894-022-01136-y

Keynote webinar | Spotlight on medication adherence

Springer Medicine

CALN1 hypomethylation as a biomarker for high-risk bladder cancer

Abstract

Background

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

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