Suppression of apoptosis and granulocyte colony-stimulating factor-induceddifferentiation by an oncogenic form of Cbl

Objective. The retroviral oncogene v-Cbl causes pre-B cell lymphomas and my eloid leukemias in mice, and its Drosophila homologue is oncogenic, causing enhanced receptor tyrosine kinase signaling, The human Cbl gene resides at 11q23. The aim of this study is to determine the effect of oncogenic Cbl o n growth-regulating responses. Materials and Methods. The oncogenic mutant of Cbl (CbI Deltal-357) was tra nsfected into factor-dependent 32Dc13 myeloid cells. Consequently, cell sur vival and differentiation were measured. Lyn, Syk, MAP kinase, and phosphat idylinositol 3 ' (PI3 ')-kinase activities, protein phosphorylation, Bcl-2 promoter activity, ubiquitination, and levels of Bcl-2, Bar, Bad, and Bcl-x (L) were determined. In addition, the effect of v-Cbl on TF-1 cell survival upon granulocyte-macrophage colony-stimulating factor withdrawal was studi ed. Results. 32Dc13 and TF-I cells expressing v-Cbl showed resistance to apopto sis upon growth factor withdrawal, and 32Dc13 cells completely failed to re spond to granulocyte colony-stimulating factor's induction of differentiati on. Basal activities of Lyn, Syk, and PI3 ' -kinase were elevated in the v- Cbl line. There was neither enhanced tyrosine phosphorylation of cellular p rotein content, Chi, or Jak2, nor serine phosphorylation of MAP kinase or A kt, After factor withdrawal, the level of Bcl-2 was greater in v-Cbl cells than in central cells, Conclusions. Neither increased Bcl-2 promoter activity nor decreased ubiqui tination of Bcl-2 could account for increased Bcl-2 levels, v-Cbl-expressin g 32Dc13 cells were resistant to differentiation, v-Cbl suppresses apoptosi s and differentiation, possibly through enhancement of Lyn, Syk, and PI3 ' -kinase activities and Bcl-2. (C) 2001 International Society for Experiment al Hematology Published by Elsevier Science Inc.