DNA Methylation Analysis of Steroid Hormone Receptor Genes
Por:
Camacho-Arroyo I., Hansberg-Pastor V., Rodríguez-Dorantes M.
Publicada:
1 ene 2014
Resumen:
Steroid hormone receptors (SHR) are important transcription factors for
regulating different physiological and pathological processes. Their
altered expression has been strongly associated to cancer progression.
Epigenetic marks such as DNA methylation have been proposed as one of
the regulatory mechanisms for SHR expression in cancer. DNA methylation
occurs at CpG dinucleotides, which form clusters known as CpG islands.
These islands are mostly observed at promoter regions of housekeeping
genes, and their aberrant methylation in cancer cells is associated with
silencing of tumor-suppressor gene expression. SHR genes are
characterized for presenting alternative promoters with different CpG
island content, which are prone to be methylated. The method of choice
for studying DNA methylation is bisulfite sequencing, since it provides
information about the methylation pattern at single-nucleotide level.
The method is based on the deamination of cytosine residues to uracil
after treatment with sodium bisulfite. The converted DNA is amplified by
a polymerase chain reaction, cloned, and sequenced. Here, we describe a
protocol for bisulfite sequencing suitable for analyzing different CpG
regions in SHR genes.
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