Hairpin-bisulfite sequencing of cells exposed to decitabine documents the process of DNA demethylation

ABSTRACT

Although the mechanism of DNA demethylating drugs has been understood for many years, the direct effect of these drugs on methylation of the complementary strands of DNA has not been formally demonstrated. By using hairpin-bisulphite sequencing, we describe the kinetics and pattern of DNA methylation following treatment of cells by the DNA methyltransferase 1 (DNMT1) inhibitor, decitabine. As expected, we demonstrate complete loss of methylation on the daughter strand following S-phase in selected densely methylated genes in synchronized Jurkat cells. Thereafter, cells showed a heterogeneous pattern of methylation reflecting replication of the unmethylated strand and restoration of methylation.

KEYWORDS:

• Demethylation
• decitabine
• dnmt1 inhibition

Disclosure statement

The authors report no conflict of interest.

Additional information

Funding

This work was supported by the Cancer Research Trust New Zealand under Grant GOT-1642-RPG and an Otago Medical School Collaborative Grant 2015.