RESUMEN
Differentiation of various leukemic cells can be induced by liganded retinoic acid receptors and protein phosphatase inhibitors. In this study, we explored the effects of okadaic acid (OA), the phosphatase inhibitor, and retinoic acid (RA) in v-myb-transformed monoblasts BM2. OA induced differentiation of BM2 monoblasts into macrophage-like cells, as documented by analyses of cell morphology, cell cycle, phagocytic activity, non-specific esterase activity, production of reactive oxygen species and expression of vimentin and Mo-1. In contrast to many other leukemic cell lines, BM2 cells do not respond to retinoic acid. However, once exposed to OA and RA simultaneously, BM2 cells differentiate along monocyte/macrophage pathway more efficiently. We conclude that RA enhances differentiation of v-myb-transformed monoblasts induced by protein phosphorylation.
Asunto(s)
Antineoplásicos/farmacología , Diferenciación Celular/efectos de los fármacos , Leucemia/metabolismo , Monocitos/efectos de los fármacos , Ácido Ocadaico/farmacología , Tretinoina/farmacología , Animales , Línea Celular Transformada , Línea Celular Tumoral , Pollos , Genes myb , Immunoblotting , Macrófagos/citología , Macrófagos/efectos de los fármacos , Monocitos/citología , Activación Transcripcional/efectos de los fármacos , TransfecciónRESUMEN
Maturation of blood cells depends on dramatic changes of expression profiles of specific genes. Although these changes have been extensively studied, their functional outcomes often remain unclear. In this study, we explored the identity and function of an unknown protein that was greatly overexpressed in v-myb-transformed BM2 monoblasts undergoing differentiation to macrophage-like cells. We identified this protein as vimentin, the intermediate filament protein. We show that an increased level of vimentin protein results from activation of the vimentin gene promoter occurring in monoblastic cells induced to differentiate by multiple agents. Furthermore, our studies reveal that the vimentin gene promoter is stimulated by Myb and Jun proteins, the key transcriptional regulators of myeloid maturation. Silencing of vimentin gene expression using siRNA markedly suppressed the ability of BM2 cells to form macrophage polykaryons active in phagocytosis and producing reactive oxygen species. Taken together, these findings document that up-regulation of vimentin gene expression is important for formation of fully active macrophage-like cells and macrophage polykaryons.