Human
breast cancer cell lines, as well as transformed mammary epithelial cells (HBL-100) and growth-stimulated normal breast epithelial cells showed positive cytochemical reaction with the
proteinase substrate 2-(N-benzyloxycarbonyl-L-arginyl-L-arginylamido)-4-methoxynapht halene, in the presence of
5-nitrosalicylaldehyde. The reaction product, small fluorescent granules, was distributed throughout the cytoplasm, in the perinuclear zone, in some cytoplasmic projections, and at the cell surface. Using a panel of various
proteinase inhibitors, we found that the formation of the reaction product was an enzymic function of a
cysteine proteinase. Using the substrate 7-(N-benzyloxycarbonyl-L-arginyl-L-arginylamido)-4-methylcoumarin, we evaluated some biochemical properties of the
cysteine proteinase, including pH-activity profile, pH stability, apparent relative molecular mass and sensitivity toward various
proteinase inhibitors. We found that the
proteinase from the studied breast epithelial cells exhibited characteristics of a mature form of
cathepsin B. Taken together, the cytochemical and biochemical data provide evidence that human breast epithelial cells of
cancer origin, as well as in the transformed or growth-stimulated state express active
cathepsin B and compartmentalize it into specific subcellular sites.