Differential expression of keratins 13 and 16 in normal epithelium, benign lesions, and ductal carcinomas of the human breast determined by the monoclonal antibody Ks8.12.

ABSTRACT

Previous electrophoretic analysis has indicated that keratins 13 and 16 (K13 and K16) are not present in normal human breast epithelium, but K16 is expressed in some ductal carcinomas (Moll et al., Cell, 31 11-24, 1982). K16 may thus represent a marker for identifying a subset of ductal carcinomas and for distinguishing such tumor cells from normal cells. To explore this possibility further, we have used the monoclonal antibody Ks8.12, reportedly specific for K13 and K16 (Huszar et al., Differentiation, 31 141-153, 1986), to examine keratin expression in human breast tissues. In immunocytochemistry the antibody reacted with epithelial cells of all normal samples, hyperplasias, and fibroadenomas. The staining was moderate to strong and always heterogeneous, involving most but not all luminal cells of ducts and acini. Myoepithelial cells were never stained. Of twenty-one ductal carcinomas examined, 90% gave a very weak or no reaction. The remaining 10% of cancers exhibited moderate to strong staining in about 50% of tumor cells. Cytoskeletal polypeptides extracted from the tissues and separated by polyacrylamide gel electrophoresis were analyzed by Western blotting to identify the polypeptides recognized by Ks8.12. In samples from normal tissue and a fibroadenoma, the antibody recognized a major Mr 48,000 component, a size appropriate for K16. In extracts from a hyperplasia and two of four carcinomas, the antibody detected a Mr 54,000 polypeptide, as well as a Mr 48,000 band, properties consistent with K13 and K16, respectively. Our results provide the first evidence indicating that K16 is present in normal as well as abnormal human breast epithelium. In addition, the data suggest that K13 may be expressed in some benign lesions and ductal carcinomas of the breast. Accordingly, Ks8.12 may prove to be useful for subclassifying ductal carcinomas and for discriminating between normal and certain benign and malignant disorders of human mammary epithelium.