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Updated: May 16, 2026
Comprehensive DNA Methylation Analysis Using a Methyl-CpG-binding Domain Capture-based Method in Chronic Lymphocytic Leukemia Patients
Published on: June 16, 2017
DNA methylation in ATRA-treated leukemia cell lines lacking a PML-RAR chromosome translocation
Regina Miftakhova1, Tove Sandberg, Andreas Hedblom
1Department of Laboratory Medicine, Skane University Hospital, Lund University, 205 02 Malmö, Sweden.
Abstract:
A deficient retinoic acid signaling has been suggested to be an important cause of the clinical inefficacy of all-trans retinoic acid (ATRA) therapy in non-promyelocytic (non-PML) forms of acute myeloid leukemia (AML). The general aim of the present work was to explore novel ways to take advantage of the anti-leukemic potential of ATRA, and, specifically, to search for a synergism between ATRA and epigenetic drugs. Because previous reports have found no major influence of ATRA on DNA methylation, we investigated whether ATRA-mediated differentiation of the U937 and HL-60 AML cell lines, both lacking a PML-retinoic acid receptor (RAR) fusion product, is accompanied by early-appearing and weak changes in CpG methylation. We report that in HL-60 cells, by using a highly quantitative analysis of a set of genes found to be abnormally expressed in AML, polymerase chain reaction (PCR)-amplified p16 gene promoter molecules (each with 15 CpG sites), exhibited a CpG methylation level of 0-4% in untreated cells, which increased to 4-21% after treatment with ATRA for seven days. In contrast to HL-60 cells, U937 cells exhibited a very high CpG methylation level in p16, and ATRA did not influence the promoter methylation of this gene. In the total CCGG sites of the genome, analysed using a methylation-sensitive restriction enzyme, CpG methylation was significantly lower in ATRA-treated HL-60 (p<0.01) and U937 cells (p<0.05) than in controls. Taken together, our findings show that ATRA can influence DNA methylation, and suggest that future research should investigate whether epigenetic modulation may evoke a clinical effect of ATRA in leukemia.
Insights
All-trans retinoic acid (ATRA) influences DNA methylation in acute myeloid leukemia (AML) cells. This suggests that combining ATRA with epigenetic drugs could improve AML treatment efficacy.
Area of Science:
- Oncology
- Molecular Biology
- Epigenetics
Background:
- All-trans retinoic acid (ATRA) therapy is often ineffective in non-promyelocytic (non-PML) acute myeloid leukemia (AML).
- Deficient retinoic acid signaling is a suspected cause of this clinical inefficacy.
- Previous research indicated ATRA has minimal impact on DNA methylation.
Purpose of the Study:
- To explore novel strategies for leveraging ATRA's anti-leukemic potential.
- To investigate potential synergistic effects between ATRA and epigenetic drugs.
- To examine if ATRA-induced differentiation in AML cell lines is associated with early changes in DNA methylation.
Main Methods:
- Utilized U937 and HL-60 AML cell lines, both lacking PML-RAR fusion.
- Quantitatively analyzed CpG methylation in the p16 gene promoter after ATRA treatment.
- Assessed global CpG methylation using methylation-sensitive restriction enzymes.
Main Results:
- In HL-60 cells, ATRA treatment increased p16 promoter CpG methylation from 0-4% to 4-21%.
- ATRA did not alter the high p16 promoter methylation in U937 cells.
- Global CpG methylation significantly decreased in ATRA-treated HL-60 and U937 cells compared to controls.
Conclusions:
- ATRA can influence DNA methylation patterns in AML cells.
- Findings suggest ATRA's effect on DNA methylation warrants further investigation.
- Epigenetic modulation might enhance the clinical efficacy of ATRA in leukemia treatment.
