Combined effects of onconase and IFN-beta on proliferation, macromolecular syntheses and expression of STAT-1 in JCA-1 cancer cells.

Prostate cancer (CaP), the most common malignancy in American men, presents its greatest challenge to clinicians when the cancer progresses to the hormone-refractory state. In the present investigation, we studied the combined effects of interferon (IFN) and onconase, each of which has reported antitumor activity, on growth and specific protein expression in JCA-1 cells. Cells were treated for up to 3 days with 1 and 5 microg/ml onconase, with and without concurrent addition of IFN-beta(ser) (10(3) IU/ml). Cell count and viability, and de novo RNA and protein synthesis were determined. Expression and subcellular distribution of STAT-1 were also assessed by immunoblot analysis. JCA-1 cells treated for 3 days with IFN or onconase showed a 15-30% reduction in cell proliferation, which was increased to 42-51% with both agents. Analysis of [35S]methionine incorporation into cells confirmed a more pronounced inhibitory effect elicited by IFN-beta and onconase; IFN-beta and 1 microg/ml onconase each decreased de novo protein synthesis by 23-25%, while the combination resulted in 59% suppression. Similar studies using incorporation of [3H]uridine into RNA yielded less significant effects. Further investigation using pre-labeled cellular RNA and proteins showed that either agent or their combination did not affect the turnover of macromolecules. To test whether the antiproliferative effects of IFN-beta and onconase were correlated with one or more specific gene changes, expression of an IFN-modulated protein, STAT-1, was determined. Both phosphorylated and unphosphorylated forms of STAT-1 and its subcellular distribution in the nucleus and cytoplasm, were increased 3-fold by IFN-beta. The IFN-elicited elevation of STAT-1 was not additionally augmented by onconase but was reduced 20-25% when onconase was simultaneously present as IFN-beta. These data show that the overall changes in STAT-1 did not correlate with the reduction in cell growth and the suppression of de novo protein/RNA synthesis elicited by these two agents, and imply that other target proteins are likely to be involved in the combined effects of IFN-beta and onconase in JCA-1 cells.