title: Promoter DNA methylation and expression levels of HOXA4, HOXA5 and MEIS1 in acute myeloid leukemia.
abstract: HOXA genes encode transcription factors, which are crucial for embryogenesis and tissue differentiation and are involved in the early stages of hematopoiesis. Aberrations in HOXA genes and their cofactor MEIS1 are found in human neoplasms, including acute myeloid leukemia (AML). The present study investigated the role of HOXA4, HOXA5 and MEIS1 promoter DNA methylation and mRNA expression in AML. Samples from 78 AML patients and 12 normal bone marrow (BM) samples were included. The levels of promoter DNA methylation were determined using quantitative methylation‑specific polymerase chain reaction (PCR; qMSP) and the relative expression levels were measured using reverse transcription quantitative PCR in Ficoll‑separated BM mononuclear cells and in fluorescent activated cell sorting‑sorted populations of normal hematopoietic progenitors. In total, 38.1 and 28.9% of the patients exhibited high methylation levels of HOXA4 and HOXA5, respectively, compared with the control samples, and MEIS1 methylation was almost absent. An inverse correlation between HOXA4 methylation and expression was identified in a group of patients with a normal karyotype (NK AML). An association between the genes was observed and correlation between the DNA methylation and expression levels of the HOXA gene promoter with the expression of MEIS1 was observed. Patients with favorable chromosomal aberrations revealed a low level of HOXA4 methylation and decreased expression levels of HOXA5 and MEIS1 compared with the NK AML and the adverse cytogenetic risk patients. The NK AML patients with NPM1 mutations exhibited elevated HOXA4 methylation and expression levels of HOXA5 and MEIS1 compared with the NPM1 wild‑type patients. Comparison of the undifferentiated BM‑derived hematopoietic CD34+CD38low, CD34+CD38+ and CD15+ cells revealed a gradual decrease in the expression levels of these three genes and an increase in HOXA4 promoter methylation. This differentiation‑associated variability was not observed in AML, which was classified according to the French‑American‑British system.
authors.affiliation: Department of Molecular and Translational Oncology, Maria Sklodowska‑Curie Memorial Cancer Center and Institute of Oncology, Warsaw 02‑781, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Musialik
authors.affiliation: Department of Molecular and Translational Oncology, Maria Sklodowska‑Curie Memorial Cancer Center and Institute of Oncology, Warsaw 02‑781, Poland.
authors.forename: Mateusz
authors.initials: M
authors.lastname: Bujko
authors.affiliation: Department of Molecular and Translational Oncology, Maria Sklodowska‑Curie Memorial Cancer Center and Institute of Oncology, Warsaw 02‑781, Poland.
authors.forename: Paulina
authors.initials: P
authors.lastname: Kober
authors.affiliation: Department of Immunology, Maria Sklodowska‑Curie Memorial Cancer Center and Institute of Oncology, Warsaw 02‑781, Poland.
authors.forename: Monika Anna
authors.initials: MA
authors.lastname: Grygorowicz
authors.affiliation: Department of Hematology, Oncology and Internal Diseases, The Medical University of Warsaw, Warsaw 02‑097, Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Libura
authors.affiliation: Department of Hematology, Oncology and Internal Diseases, The Medical University of Warsaw, Warsaw 02‑097, Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Przestrzelska
authors.affiliation: Department of Diagnostic Hematology, Institute of Hematology and Transfusion Medicine, Warsaw 02‑109, Poland.
authors.forename: Przemysław
authors.initials: P
authors.lastname: Juszczyński
authors.affiliation: Department of Diagnostic Hematology, Institute of Hematology and Transfusion Medicine, Warsaw 02‑109, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Borg
authors.affiliation: Department of Hematology, Jagiellonian University, Cracow 31‑501, Poland.
authors.forename: Izabela
authors.initials: I
authors.lastname: Florek
authors.affiliation: Department of Hematology, Jagiellonian University, Cracow 31‑501, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Jakóbczyk
authors.affiliation: Department of Molecular and Translational Oncology, Maria Sklodowska‑Curie Memorial Cancer Center and Institute of Oncology, Warsaw 02‑781, Poland.
authors.forename: Janusz Aleksander
authors.initials: JA
authors.lastname: Siedlecki
mesh_terms: D000328:Adult; D000368:Aged; D001854:Bone Marrow Cells / Q000378:metabolism / Q000473:pathology; D016022:Case-Control Studies; D002869:Chromosome Aberrations; D019175:DNA Methylation; D005260:Female; D015973:Gene Expression Regulation, Leukemic; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009154:Mutation; D000075064:Myeloid Ecotropic Viral Integration Site 1 Protein; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics; D000090243:Nucleophosmin; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D014157:Transcription Factors; D051941:fms-Like Tyrosine Kinase 3 / Q000235:genetics
chemical_list: C489213:HOXA5 protein, human; D018398:Homeodomain Proteins; C000616391:MEIS1 protein, human; D000075064:Myeloid Ecotropic Viral Integration Site 1 Protein; C000716967:NPM1 protein, human; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D000090243:Nucleophosmin; C073547:HOXA4 protein, human; D051941:fms-Like Tyrosine Kinase 3