We generated anti-human E1-STS monoclonal antibodies to localize estrone sulfatase (E1-STS) in human breast carcinomas. In particular, we examined the MCF-7 clone E3, ZR-75-1, MDA-MB 231, and MDA-MB-468 breast cancer cell lines and 25 breast carcinomas by either immunohistochemistry or Western blotting analysis. Simultaneously, we analyzed histological data, estrogen receptor (ER) status, progesterone receptor (PgR) status and epidermal growth factor receptor (EGFR) in breast tissue. All were surgical specimens from female patients. Nine of 25 carcinomas were obtained from premenopausal women, and 16 carcinomas were obtained frompostmenopausal women. All cell lines demonstrated positive staining for E1-STS.Interestingly, fine granulated staining of E1-STS on the cell membrane was observed. In addition, Western blotting analysis detected a 65 kD protein with an E1-STS specific band in all breast cancer cell lines regardless of the presence orabsence of E2. Twenty-two of 25 (88.0%) carcinomas showed positive staining forE1-STS, whereas negative staining was observed in the interstitial tissue surrounding tumors. In the premenopausal patients, 8 of 10 carcinomas (80.0%) showed positive staining for E1-STS, whereas 14 of 15 carcinomas (93.3%) revealed positive staining in the postmenopausal patients. The frequency of E1-STS expression was relatively higher in postmenopausal patients than in premenopausal patients but not statistically significant. The intensity of immunostaining for E1-STS depended upon the size of the tumor (NS). There was no correlation between E1-STS expression and other parameters. This evidence suggests E1-STS expression may beinvolved in the development of breast cancer. Further studies are necessary to clarify the relationship between E1-STS expression and prognostic factors. Immunoreactive E1-STS may be localized in cancer cells but not in surrounding tissuesin breast cancer.