Expression of the imprinted tumour-suppressor gene H19 is tightly regulated during normal haematopoiesis and is reduced in haematopoietic precursors of patients with the myeloproliferative disease polycythaemia vera

• Published in: The Journal of pathology 2000-01, Vol.190 (1), p.61-68
• Main Authors: Núnêz, César, Bashein, Abdulla M., Brunet, Clare L., Hoyland, Judith A., Freemont, Anthony J., Buckle, Anne-Marie, Murphy, Clare, Cross, Michael A., Lucas, Guy, Bostock, Victoria J., Brady, Gerard
• Format: Article
• Language: English
• Subjects: Animals; Antigens, CD34; Biological and medical sciences; Flow Cytometry; Gene Expression; Genes, Tumor Suppressor; H19; H19 gene; Hematologic and hematopoietic diseases; Hematopoiesis - genetics; Hematopoietic Stem Cells - metabolism; Humans; IGF2; In Situ Hybridization; Insulin-Like Growth Factor II - genetics; leukaemia; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis; lineage commitment; Medical sciences; Mice; Muscle Proteins - genetics; Muscle Proteins - metabolism; polycythemia vera; Polycythemia Vera - genetics; Polycythemia Vera - immunology; Reverse Transcriptase Polymerase Chain Reaction; RNA, Long Noncoding; RNA, Untranslated; RT-PCR; tumour suppressor
• ISSN: 0022-3417; 1096-9896
• DOI: 10.1002/(SICI)1096-9896(200001)190:1<61::AID-PATH502>3.0.CO;2-#

cDNA subtraction was employed to uncover differences in gene expression between myeloproliferative polycythaemia vera (PV) and normal haematopoietic precursors. Following cDNA subtraction using mRNAs isolated from PV and normal CD34+/CD33− bone‐marrow cells, expression of the tumour suppressor H19 was found to be low or absent in the PV sample. Low levels of H19 expression in PV patients were confirmed by in situ hybridization. Using semi‐quantitative reverse transcriptase‐polymerase chain reaction (RT‐PCR) to examine expression in the pluripotent haematopoietic cell line FDCP‐mix and single bone‐marrow precursors, unambiguous IGF2 and H19 expression was demonstrated in normal haematopoietic precursors. Examination of individual bone‐marrow precursors revealed that all IGF2‐expressing haematopoietic precursors also co‐expressed H19, indicating that H19 and IGF2 may be co‐ordinately regulated during haematopoiesis. Analysis of FDCP‐mix undergoing differentiation and single pluripotent and committed bone‐marrow precursors revealed that the pattern of H19 expression coincided with the commitment to a single lineage. Taken together, these observations demonstrate that H19 and IGF2 are specifically expressed during haematopoiesis and that low levels of H19 expression are associated with PV and may contribute to the pathology of the disease. Copyright © 2000 John Wiley & Sons, Ltd.