Abnormal expression of homeobox genes may lead to the development of leukemias, lymphomas, and solid tumors. Expression of homeobox genes in mammary glands, however, has not been studied actively until recently. We have examined the expression of POU homeobox genes in human breast cancer cell lines and human breast tissue samples. Using a pair of degenerate primers for reverse transcription‐polymerase chain reaction (RT‐PCR) followed by DNA sequencing, we found that the human breast cancer cell line, MCF7, expresses at least 4 POU gene products: OCT1, OCT2, OCT3 and OCT11 (Skn‐1a/i, Epoc‐1). The expression of OCT1 and OCT2 in other human breast epithelial cell lines was further determined by Western blot analyses and electrophoretic mobility shift assay. We were unable to detect OCT11 in human breast cancer cell lines using the anti rat Skn‐1a/i antibody, although the expression of this gene in both human breast cancer cell lines and human primary breast tumors was detected by RT‐PCR. OCT3 is an embryonic transcription factor. We found that this gene is also expressed in human breast cancer cell lines and all human primary breast carcinomas examined, but not in normal human breast tissue. Taken together, we have shown that several POU genes are expressed in human breast epithelial cells. As OCT3 expression was detected only in the breast cancerous cells, this embryonic transcription factor could play an important role in mammary gland carcinogenesis. Int. J. Cancer 81:104–112, 1999. © 1999 Wiley‐Liss, Inc.