Abstract
Epigenetic gene regulation is important in human cancer. Both functional and observational data implicate alterations of histone modifications, DNA promoter methylation, and non-coding RNA expression in carcinogenic roles. We sought to explore the role of aberrant DNA hypermethylation in the regulation of microRNA (miR) expression in human cancer. From human genome databases we calculated that 13 and 28% of human miR genes are located within 3 and 10 kb of a CpG island, respectively. To identify miRs that are regulated by epigenetic mechanisms in cancer, we performed expression profiling prior to and following treatment of cell lines with 5-azacytidine. We used oligonucleotide microarrays to determine miR expression. For miRs whose expression changed following 5-azacytidine treatment, we sequenced the adjacent CpG island and promoter using bisulphite-treated DNA. Here, we describe these methods to enable other researchers to use this approach.
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Acknowledgments
The authors would like to thank Mark Meuth, Freddie Hamdy, Helen Owen, Saiful Miah, Helen Bryant, Katie Myers, Maggie Glover, Ewa Dudziec, Louise Goodwin, Leila Ayandi, and Sheila Blizard for their help, support, and encouragement with this work. This work was funded by a GSK Clinician Scientist fellowship to JWFC and project grants to JWFC from Yorkshire Cancer Research, Sheffield Hospitals Charitable trust, and the European Union (Framework 7).
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Choudhry, H., Catto, J.W.F. (2011). Epigenetic Regulation of MicroRNA Expression in Cancer. In: Wu, W. (eds) MicroRNA and Cancer. Methods in Molecular Biology, vol 676. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-863-8_12
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DOI: https://doi.org/10.1007/978-1-60761-863-8_12
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Publisher Name: Humana Press, Totowa, NJ
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