Analysis of mechanism of human megakaryocyte maturation

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The mechanism of maturation of human megakaryocytic cells has been investigated.1. The role of Evi-1 on megakaryocytic cells was investigated. Evi-1 m-RNA was expressed in two megakaryoblastic leukemia cell lines, CMK and Meg-J.However Evi-1 m-RNA expression did not increase after stimulation by TPA (12-0-tetradecanoyl phorbol-13-acetate), IL-3 (interleukin-3), IL-6 or IL-11. Myb m-RNA was expressed in CMK cells however, addition of oligonucleotide of myb to CMK cells chould not suppress the proliferation, suggesting that myb has no role for the proliferation of CMK cells.2. When Meg-J cells were cultured in the presence of K252a, a protein kinase C inhibitor, the ploidy was dramatically shifted toward higher values. This assay system may be useful for studying the mechnism of polyploidization of megakaryocytic cells.3. Enhancement of CD41 expression of CMK cells by TPA was ablogated by Herbimycin A,a tyrosine kinase inhibitor, indicating that TPA-induced CD41 expression was mediated by tyrosine kinases.