Establishment and characterization of an immature human megakaryoblastic cell line, MEG-A2.
We have established a novel human megakaryoblastic cell line, designated as MEG-A2, from a patient with megakaryoblastic crisis of Philadelphia (Ph) chromosome positive chronic myelogenous leukemia. MEG-A2 cells showed positive phenotypes for periodic acid Schiff and alpha-naphthylbutyrate esterase reactions, but were negative for myeloperoxidase and naphthol ASD chloroacetate esterase reactions. Flow cytometric analyses of cell surface markers revealed that MEG-A2 cells had a low level of GP IIb/IIIa expression as well as apparent expressions of CD4, CD7, CD13, CD33 and CD34 antigens, but no expression of GP Ib nor glycophorin A. Stimulation with phorbol 12-myristate 13-acetate (PMA) dramatically increased the expression of megakaryocyte-related markers such as HPL-3, J15, Pit-1, Y2/51 and AN51 in MEG-A2 cells. The PMA-stimulation also induced expression of platelet peroxidase (PPO) in MEG-A2 cells on electromicroscopic observation. Proliferative responses to granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3) or erythropoietin were observed, and the expression of GP IIb/IIIa was increased by stimulation with GM-CSF, IL-3, erythropoietin and interleukin-6 (IL-6). Protein S mRNA expression was seen in cultured cells on Northern blot analysis. Expression of platelet factor 4 mRNA was induced in PMA-stimulated cells, and a marked accumulation of protein was observed in the culture medium. In conclusion, a new cell line, MEG-A2, belongs to the relatively immature megakaryocytic lineage and has markedly increased megakaryocytic characteristics with PMA stimulation.
Abe
A
A
First Department of Internal Medicine, Nagoya University School of Medicine, Japan.
Emi
N
N
Kato
H
H
Adachi
K
K
Murate
T
T
Saga
S
S
Ogura
M
M
Kojima
T
T
Tanimoto
M
M
Morishita
N
N
D000328:Adult; D000782:Aneuploidy; D015703:Antigens, CD / Q000032:analysis; D000943:Antigens, Differentiation / Q000032:analysis; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000032:analysis; D001752:Blast Crisis / Q000473:pathology; D002265:Carboxylic Ester Hydrolases; D002455:Cell Division / Q000187:drug effects; D017809:Fatal Outcome; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000187:drug effects / Q000473:pathology*; D006801:Humans; D007621:Karyotyping; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics / Q000473:pathology*; D008297:Male; D008533:Megakaryocytes; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000032:analysis; D014411:Neoplastic Stem Cells / Q000737:chemistry / Q000187:drug effects / Q000473:pathology*; D010978:Platelet Factor 4 / Q000096:biosynthesis / Q000235:genetics; D010980:Platelet Membrane Glycoproteins / Q000032:analysis; D017293:Protein S / Q000096:biosynthesis / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics; D018121:Receptors, Cytokine; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000235:genetics; D053628:Receptors, Thrombopoietin; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000187:drug effects; D001620:beta-Thromboglobulin / Q000096:biosynthesis / Q000235:genetics
D015703:Antigens, CD; D000943:Antigens, Differentiation; D014408:Biomarkers, Tumor; D016298:Hematopoietic Cell Growth Factors; D009363:Neoplasm Proteins; D010980:Platelet Membrane Glycoproteins; D017293:Protein S; D011518:Proto-Oncogene Proteins; D018121:Receptors, Cytokine; D011971:Receptors, Immunologic; D053628:Receptors, Thrombopoietin; D001620:beta-Thromboglobulin; C082218:MPL protein, human; D010978:Platelet Factor 4; D002265:Carboxylic Ester Hydrolases; C021855:naphthylbutyrate esterase; D013755:Tetradecanoylphorbol Acetate