INTERNALIZATION OF THE BOWMAN-BIRK PROTEASE INHIBITOR BY INTESTINAL EPITHELIAL-CELLS

Protease inhibitors have been shown to be effective suppressors of carcinogenesis in vitro and in vivo. For example, the soybean-derived Bowman-Birk inhibitor (BBI) suppresses dimethylhydrazine-induced colon carcinogenesis in mice. Relatively little is known about the effects of protease inhibitors on intestinal epithelial cells. In the present study, we have investigated the interaction of the anticarcinogenic BBI with intestinal epithelial cells. At the concentrations examined, BBI was non-toxic and had no effect on the doubling time, saturation density or rate of DNA synthesis by these cells. This compound was taken up by these cells in a time dependent manner and was present in the cells for 12 h following a 2 h incubation with BBI. Subcellular fractionation experiments demonstrated that the bulk of the internalised inhibitor was present in the cytosol. Analysis of BBI from treated cells on a chymotrypsin affinity column revealed that active inhibitor was present in the cells. Our results indicate that the BBI is internalised by colonic epithelial cells which would allow BBI to inhibit critical intracellular proteases and thus suppress malignant transformation.