Aberrant methylation of the M-type phospholipase A2 receptor gene in leukemic cells
1 Institut für Klinische Chemie und Laboratoriumsmedizin, Technische Universität Dresden, Fetscherstrasse 74, D-01307, Dresden, Germany
2 Medizinische Klinik und Poliklinik I, Technische Universität Dresden, Dresden, Germany
BMC Cancer 2012, 12:576 doi:10.1186/1471-2407-12-576Published: 5 December 2012
Additional file 1:
Table S1. Characteristics of patients analyzed for PLA2R1 methylation using MS-HRM analysis of bisulfite-modified genomic DNA.
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Additional file 2:
Table S2. Characteristics of MDS patients with different IPSS classifications. The degree of PLA2R1 methylation shown was measured using MS-HRM analysis of bisulfite-modified genomic DNA from bone marrow aspirates. CMML-1, chronic myelomonocytic leukemia; RCMD, refractory cytopenia with multilineage dysplasia; RCMD-RS, refractory cytopenia with multilineage dysplasia and ringed sideroblasts; RAEB, refractory anemia with excess blasts; RCUD, refractory cytopenia with unilineage dysplasia; RAEB-t, refractory anemia with excess blasts in transformation; sAML, secondary acute myeloid leukemia.
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Additional file 3:
Table S3. Characteristics of patients treated with azacitidine. The degree of PLA2R1 methylation shown was measured using MS-HRM analysis of bisulfite-modified genomic DNA from blood samples. RCMD, refractory cytopenia with multilineage dysplasia; RAEB, refractory anemia with excess blasts; RAEB-t, refractory anemia with excess blasts in transformation.
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Additional file 4:
Figure S1. Melt curve analyses of bisulfite-modified and amplified PLA2R1 gene sequences in bone marrow aspirates of MDS patients. After isolation and subsequent bisulfite modification of genomic DNA from bone marrow aspirates melt properties of amplified PLA2R1 sequences covering 5`-CpG sites 6–14 (−437 bp to −270 bp from exon 1) were analyzed using RotorGene Q. P34; a 74 years old male patient with RCMD and low-risk according to IPSS classification, P57; a 69 years old male patient with RAEB-2 and intermediate-1-risk, P66; a 69 years old female patient with RAEB-2 and intermediate-2-risk, and P69; a 56 years old male patient with AML and high-risk classification. Individual data of MDS patients are summarized in Additional file 2: Table S2. Melt curves of amplified genomic DNA from MDS patients (in colors) and those of unmethylated (0%) and methylated (100%) standard DNA samples (in black) are shown. Fluorescence dF/dT were measured. Analyses were performed in duplicates and results are representative of three independent measurements.
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Additional file 5:
Table S4. MS-HRM analyses of bisulfite-modified genomic DNA isolated from peripheral blood samples of a 58 years old female patient with minimal residual disease. After treatment of AML, FAB M2, by allogeneic hematopoietic stem cell transplantation three years ago the patient underwent a pre-emptive treatment with azacitidine to avoid haematological relapse. Amounts of leukocytes, blasts, and PLA2R1 methylation degrees during treatment measured using MS-HRM analysis are summerized. n; normal melt curve without methylated DNA fraction, p; pathological melt curve with methylated DNA fraction in addition to unmethylated DNA fraction.
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