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A Versatile Assay for Detection of Aberrant DNA Methylation in Bladder Cancer

  • Stella TommasiEmail author
  • Ahmad Besaratinia
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1655)

Abstract

Urothelial carcinoma of the bladder is one of the most common malignancies in the industrialized world, mainly caused by smoking and occupational exposure to chemicals. The favorable prognosis of early stage bladder cancer underscores the importance of early detection for the treatment of this disease. The high recurrence rate of this malignancy also highlights the need for close post-diagnosis monitoring of bladder cancer patients. As for other malignancies, aberrant DNA methylation has been shown to play a crucial role in the initiation and progression of bladder cancer, and thus holds great promise as a diagnostic and prognostic biological marker. Here, we describe a protocol for a versatile DNA methylation enrichment method, the Methylated CpG Island Recovery Assay (MIRA), which enables analysis of the DNA methylation status in individual genes or across the entire genome. MIRA is based on the ability of the methyl-binding domain (MBD) proteins, the MBD2B/MBD3L1 complex, to specifically bind methylated CpG dinucleotides. This easy-to-perform method can be used to analyze the methylome of bladder cancer or urothelial cells shed in the urine to elucidate the evolution of bladder carcinogenesis and/or identify epigenetic signatures of chemicals known to cause this malignancy.

Key words

Aromatic amines Biomarkers Epigenetics Methyl-binding domain (MBD) proteins Tobacco smoke Urine 

Notes

Acknowledgments

ST and AB declare no conflicts of interest.

Work of the authors is funded by grants from the National Institute of Dental and Craniofacial Research of the National Institutes of Health (1R01DE026043-01) to AB and from the University of California Tobacco-Related Disease Research Program (TRDRP-25IP-0001) to ST.

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  1. 1.Department of Preventive Medicine, USC Keck School of MedicineUniversity of Southern CaliforniaLos AngelesUSA

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