Four human cell lines derived from Ewing's sarcoma, EW-7, EW-1, COH and ORS, were investigated to establish the eects of human recombinant interferon-a2a and human recombinant interferon-b on cell proliferation and apoptosis. All four cell lines were much more sensitive to the antiproliferative eects of IFN-b than of IFN-a. Analysis of the early signals triggered by IFN-a and IFN-b demonstrated that the two IFNs were similarly eective in inducing tyrosine phosphorylation of the Jak-1 and Tyk-2 kinases and the transcription factors Stat-1 and Stat-2. Interestingly, an additional rapid phosphorylation of Stat-1 on serine was observed after IFN-b treatment, with concomitant activation of p38 mitogen-activated protein kinase. In these cells, Stat-1 Ser727 phosphorylation in response to IFN-b was found to be impaired by p38 MAPkinase inhibitor (SB203580). IFN-b induced the formation of the Interferon Stimulated Gene Factor 3 complex more eciently than IFN-a, as well as sustained induction of IRF-1, which may account for its greater induction of 2'5'oligo(A)synthetase and greater inhibition of cell proliferation. IFN-b, but not IFN-a, induced apoptosis in wild-type p53 EW-7 and COH cell lines, but not in the mutated p53 EW-1 or ORS cell lines. The apoptosis induced by IFN-b in EW-7 and COH cell lines appeared to be mediated by IRF-1 and involved the activation of caspase-7. Ectopic expression of IRF-1 induced apoptosis in all four cell lines which correlated with the activation of caspase-7 and with the downregulation of the Bcl-2 oncoprotein, as observed for IFN-b-induced apoptosis in parental EW-7 and COH cell lines. Oncogene (2000) 19, 3372 ± 3383.