Epigenetic changes in FOXO3 and CHEK2 genes and their correlation with clinicopathological findings in myelodysplastic syndromes

Hematol Oncol Stem Cell Ther. 2020 Dec;13(4):214-219. doi: 10.1016/j.hemonc.2019.11.004. Epub 2020 Mar 18.

Abstract

Objectives/background: Myelodysplastic syndromes (MDSs) are a heterogeneous disease in terms of clinical course and response to therapy. Epigenetic changes are the primary mechanism of MDS pathogenesis. FOXO3 and CHEK2 genes play significant roles in normal cellular mechanisms and are also known as tumor suppressor genes. We aimed to clarify the correlation of epigenetic changes in these genes with clinicopathologic findings in MDS.

Methods: A total of 54 newly diagnosed MDS patients referred to Shariati and Firouzgar Hospitals (Tehran, Iran) were included in the study from 2013 to 2015, comprising the following cases: 26 with refractory cytopenia with unilineage dysplasia, 10 with refractory cytopenia with multilineage dysplasia, four refractory anemia with excess blasts-1 (RAEB-1), 11 refractory anemia with excess blasts-2 (RAEB-2), and three MDS associated with isolated deletion (5q-). Risk groups were determined according to the Revised International Prognostic Scoring System (IPSS-R). The methylation status of CHEK2 and FOXO3 promoters were determined by methylation-sensitive high-resolution melting analysis of sodium bisulfite-converted DNA. Expressions of CHEK2, FOXO3, and GAPDH were measured by quantitative real-time polymerase chain reaction and fold changes were calculated using the ΔΔCT method.

Results: Statistical analysis revealed no promoter methylation of CHEK2 and FOXO3 in healthy control specimens. FOXO3 promoter methylation was associated with high-risk World Health Organization subgroups (p = .017), high-risk IPSS-R (p = .007), high-risk cytogenetics (p = .045), and more than 5% blasts in bone marrow (p = .001). CHEK2 promoter methylation was correlated with more than 5% blasts in bone marrow (p = .009).

Conclusions: Promoter methylation of CHEK2 and especially FOXO3 is associated with adverse clinicopathological findings and disease progression in MDS.

Keywords: DNA hypermethylation; Epigenetic changes; High-resolution melting; Myelodysplastic syndromes; Real-time PCR.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Checkpoint Kinase 2* / genetics
  • Checkpoint Kinase 2* / metabolism
  • DNA Methylation*
  • Epigenesis, Genetic*
  • Female
  • Forkhead Box Protein O3* / genetics
  • Forkhead Box Protein O3* / metabolism
  • Humans
  • Male
  • Middle Aged
  • Myelodysplastic Syndromes* / genetics
  • Myelodysplastic Syndromes* / metabolism
  • Promoter Regions, Genetic*

Substances

  • FOXO3 protein, human
  • Forkhead Box Protein O3
  • Checkpoint Kinase 2
  • CHEK2 protein, human