title: Differentiation-induced changes in protein-tyrosine phosphatase activity and commensurate expression of CD45 in human leukemia cell lines.
abstract: Pharmacologic differentiation of the promyelocytic leukemia HL60 is associated with an increase in cellular tyrosine phosphatase activity. We asked (a) if this increase might, at least in part, be due to changes in a transmembranous protein-tyrosine phosphatase, CD45; and (b) if CD45 changes similarly in other differentiating leukemias. Differentiation of HL60, several chronic myelogenous leukemias, a monocytic leukemia (THP-1), and a monoblastoid leukemia (U-937) could be induced by phorbol ester, 1,25-dihydroxy vitamin D3, dimethyl sulfoxide, or cyclic AMP analogues. This differentiation was associated with a marked increase in (a) total cellular tyrosine phosphatase activity (2-4-fold as measured by the ability to dephosphorylate a tyrosine-phosphorylated peptide); (b) CD45-specific tyrosine phosphatase activity (2-4-fold); (c) CD45 cell surface expression by flow cytometry (2-5-fold); (d) synthesis of both exon B-dependent M(r) 205,000 and exon ABC- M(r) 185,000 CD45 proteins, as revealed by immunoprecipitation with antisera specific for CD45 isoforms. Both isoforms have enhanced electrophoretic mobility when isolated from the differentiated cells. This enhanced mobility did not appear to be due to decreased stoichiometry of CD45 phosphorylation on serine/threonine residues. Interestingly, 12-O-tetradecanoylphorbol-13-acetate transiently reduced CD45 protein-tyrosine phosphatase activity in the chronic myelogenous leukemia cell RWLeu4 without altering the CD45 amount (as measured by cell surface immunofluorescence). Modulation of CD45 tyrosine phosphatase activity (and protein levels) may play a role in differentiation or in maintaining cells in a nonproliferative state or may represent a phenotypic marker of differentiation.
authors.affiliation: Department of Medicine, Roger Williams Cancer Center, Providence, Rhode Island.
authors.forename: M
authors.initials: M
authors.lastname: Buzzi
authors.forename: L
authors.initials: L
authors.lastname: Lu
authors.forename: A J
authors.initials: AJ
authors.lastname: Lombardi
authors.forename: M R
authors.initials: MR
authors.lastname: Posner
authors.forename: D L
authors.initials: DL
authors.lastname: Brautigan
authors.forename: L D
authors.initials: LD
authors.lastname: Fast
authors.forename: A R
authors.initials: AR
authors.lastname: Frackelton
mesh_terms: D015703:Antigens, CD / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D004121:Dimethyl Sulfoxide; D004586:Electrophoresis; D006649:Histocompatibility Antigens / Q000378:metabolism*; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000201:enzymology* / Q000473:pathology; D015473:Leukemia, Promyelocytic, Acute / Q000201:enzymology* / Q000473:pathology; D017493:Leukocyte Common Antigens; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate; D014407:Tumor Cells, Cultured
chemical_list: D015703:Antigens, CD; D006649:Histocompatibility Antigens; D017493:Leukocyte Common Antigens; D017027:Protein Tyrosine Phosphatases; D013755:Tetradecanoylphorbol Acetate; D004121:Dimethyl Sulfoxide