Korean J Hematol.  2012 Sep;47(3):186-193. 10.5045/kjh.2012.47.3.186.

Identification of genes underlying different methylation profiles in refractory anemia with excess blast and refractory cytopenia with multilineage dysplasia in myelodysplastic syndrome

• Lee S
• Kwon HC
• Kim SH
• Oh SY
• Lee JH
• Lee YS
• Seo D
• Han JY
• Kim HJ

• KMID: 2251952
• DOI: http://doi.org/10.5045/kjh.2012.47.3.186

Abstract

BACKGROUND
Myelodysplastic syndrome (MDS) is a preleukemic condition that transforms into acute myeloid leukemia. However, the genetic events underlying this transformation remain poorly understood. Aberrant DNA methylation may play a causative role in the disease and its prognosis. Thus, we compared the DNA methylation profiles in refractory anemia with excess blast (RAEB) to those in refractory cytopenia with multilineage dysplasia (RCMD).
METHODS
Bone marrow samples were collected from 20 patients with primary MDS (9 with RAEB and 11 with RCMD), and peripheral blood samples were collected from 4 healthy controls. These samples were assessed using a commercial whole genome-wide methylation assay. Methylation-specific polymerase chain reaction (PCR) was used to detect the methylation of candidate gene promoters in RAEB and RCMD.
RESULTS
Microarray data revealed significant hypermethylation in 69 genes within RAEB but not RCMD. Candidate genes were mapped to 5 different networks, and network 1 had the highest score due to its involvement in gene expression, cancer, and cell cycle. Five genes (GSTM5, BIK, CENPH, RERG, and ANGPTL2) were associated with malignant disease progression. Among them, the methylated promoter pairs of GSTM5 (55.5% and 20%), BIK (20% and 0%), and ANGPTL2 (44.4% and 10%) were observed more frequently in RAEB.
CONCLUSION
DNA methylation of GSTM5, BIK, and ANGPTL2 may induce epigenetic silencing and contribute to the increasing blasts and resulting MDS progression; however, the functions of these genes were not determined. Further study focusing on epigenetic silencing using various detection modalities is required.

Keyword

Myelodysplastic syndrome; DNA methylation; GSTM5; ANGPTL2; BIK

MeSH Terms

Anemia, Refractory
Anemia, Refractory, with Excess of Blasts
Bone Marrow
Cell Cycle
Disease Progression
DNA Methylation
Epigenomics
Gene Expression
Humans
Leukemia, Myeloid, Acute
Methylation
Myelodysplastic Syndromes
Polymerase Chain Reaction
Prognosis

Figure

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  Fig. 1 Cluster pattern for all microarray gene data. Abbreviations: RCMD, refractory cytopenia with multilineage dysplasia; RAEB, refractory anemia with excess blast.

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  Fig. 2 Hierarchical clustering analysis. No noteworthy hypermethylated gene cluster patterns were identified.

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  Fig. 3 Network 1 gene expression, cancer, and cell cycle. The 16 candidate genes are denoted in red and burrowed with white genes to form a disease network construct.

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  Fig. 4 Methylation-specific PCR results on GSTM5, ANGPTL2, BIK, and RERG genes from bone marrow samples. Patients were designated numbers 1-10 (RCMD) and 11-19 (RAEB). The PCR products were analyzed by electrophoresis on a 2% agarose gel. Abbreviations: N, normal peripheral blood; H, Human leukemic HL-60 cells; M, amplified products used as primers for the methylated sequence; U, amplified products used as primers for the unmethylated sequence.

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