Tissue Content of Hydroxyestrogens in Relation to Survival of Breast Cancer Patients

Purpose: The main goal of our study was to assess estrogen contents of breast tumor tissues, having different estrogen receptor status, in relation to long-term follow-up of patients. Experimental Design: Twenty-one breast cancer cases, all collected from January 1986 to January 1988 at the M. Ascoli Cancer Hospital Centre in Palermo, were included in the study and compared with 6 healthy women as a control group. Average follow-up time of patients was 144 ± 10 months. The estrogen receptor status of tissues was determined by both ligand binding and immunohistochemical assays. A high performance liquid chromatography-based approach, jointly with gas chromatography/mass spectrometry, was used to identify and measure main estrogens, various hydroxyestrogens, and their methoxy derivatives in both normal and tumor tissues. Results: Although variable concentrations of hydroxylated estrogens were detected, they consistently accounted for >80% of all of the estrogens. Significantly greater amounts of both 2- and 4-hydroxyestradiol, along with a marked increase of 16α-hydroxyestrone (OHE1), were observed in cancer with respect to normal breast tissues. A significant positive association was observed with elevated 16αOHE1 ( P = 0.015) in patients alive, leading to significantly lower ( P = 0.043) 2OHE1:16αOHE1 ratio values. Conversely, ratio values of 4:2 hydroxy+methoxy estrogens was significantly lower ( P = 0.006) in deceased patients. Using cutoff values of 1.2 for 4:2 hydroxy+methoxy ratio and 150 fmol/mg tissue for 16αOHE1 we achieved a clear-cut separation of patients, with over-cutoff patients having 147 months and under cutoff patients showing only 47 months median survival time ( P = 0.00008). Conclusions: Our data imply that individual hydroxyestrogens may have a distinct role in the onset and the clinical progression of breast cancer, with greater 16αOHE1 levels being in turn associated to cancer with respect to normal tissues and to a prolonged survival of breast cancer patients.