Polycythemia vera. V. Enhanced proliferation and phosphorylation due to vanadate are diminished in polycythemia vera erythroid progenitor cells: a possible defect of phosphatase activity in polycythemia vera.
Erythropoietin (EP) and stem cell factor (SCF) are essential growth factors for erythroid progenitor cell proliferation and differentiation in serum-free culture. It has been previously shown that burst-forming units-erythroid and colony-forming units-erythroid from patients with polycythemia vera (PV) have enhanced sensitivity to EP and SCF compared with normal erythroid progenitors, but little is known about the mechanism for this difference. In the present investigation, the effect of EP and SCF on protein tyrosine phosphorylation in day-8 normal and PV erythroid colony-forming cells, which give rise to colonies of 2-49 hemoglobinized cells, was studied. EP rapidly induced tyrosine phosphorylation of the EP receptor, whereas the most prominent phosphorylated protein induced by SCF was identified as the SCF receptor. No additional phosphorylated proteins were evident when PV cells were compared with normal cells. Culture of normal erythroid progenitors with orthovanadate, an inhibitor of protein tyrosine phosphatases, resulted in an increased number of erythroid colonies and enhanced protein tyrosine phosphorylation. However, in contrast, little enhancement was evident with PV cells. These results indicate that, although vanadate may be acting in normal erythroid progenitors as a phosphatase inhibitor that potentiates the kinase activity induced by SCF and EP, this function is diminished in PV cells. Because erythropoiesis is regulated by a balance between protein tyrosine kinase activity and protein tyrosine phosphatase activity, PV patients may have an abnormal phosphatase activity allowing increased cell proliferation.
Dai
C H
CH
Department of Medicine, Department of Veterans Affairs Medical Center, Vanderbilt University School of Medicine, Nashville, TN 37232-6305, USA.
Krantz
S B
SB
Sawyer
S T
ST
D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D016895:Culture Media, Serum-Free; D015672:Erythroid Precursor Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D004921:Erythropoietin / Q000494:pharmacology; D006801:Humans; D010766:Phosphorylation / Q000187:drug effects; D011087:Polycythemia Vera / Q000201:enzymology / Q000473:pathology*; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors / Q000172:deficiency* / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D019089:Stem Cell Factor / Q000494:pharmacology; D014638:Vanadates / Q000494:pharmacology*
D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D004921:Erythropoietin; D014638:Vanadates; D011505:Protein-Tyrosine Kinases; D019009:Proto-Oncogene Proteins c-kit; D017027:Protein Tyrosine Phosphatases