Bryostatin I is a natural product currently under clinical evaluation as an antitumor agent. Like the tumor-promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) it activates protein kinase C (PKC). Bryostatin I inhibits the growth of the human-derived A549 lung and MCF-7 adenocarcinoma cell lines, but much more weakly than TPA. The hypotheses were tested that differences between cell lines in their response to bryostatin I are related to cellular PKC isotype content, and that differences between TPA and bryostatin I in their effects on cell growth are associated with differential abilities to modulate specific PKC isoenzymes. PKC isozyme profiles were studied by Western-blot analysis in the cytosol, particulate and nuclear fractions of A549 and MCF-7 cells. PKCs-alpha, -epsilon and -zeta were detected in both cell types with predominant location in the cytosol. Separation of cytosolic PKC isoenzymes in A549 cells by hydroxylapatite column chromatography and determination of PKC activity in fractions yielded a major peak which contained PKC-alpha. Exposure of cells to bryostatin I or TPA for 30 min caused the redistribution of PKCs-alpha and -epsilon from the cytosol to the particulate and nuclear fractions in a concentration-dependent fashion. PKC-epsilon was completely down-regulated by exposure to 10 nM bryostatin I for 18 hr or to TPA for 24 hr. Down-regulation of PKC-alpha was partial at 10 nM and complete at 1 microM of either agent. Bryostatin I inhibited incorporation of [3H]-labelled thymidine into cells only transiently, whereas TPA arrested growth for several days in A549 cells and irreversibly in MCF-7 cells. A549 cells, in which PKC was depleted by exposure to phorbol ester for 9 weeks, were resistant towards bryostatin-induced inhibition of DNA synthesis. The results suggest that the susceptibility of adenocarcinoma cells towards bryostatin-induced growth delay are determined by cellular levels of PKCs-alpha and/or -epsilon. However, differences between bryostatin I and TPA in their abilities to inhibit cell growth do not seem to be intrinsically related to differences in redistribution or down-regulation of specific PKC isoenzymes.