Comparative studies of aromatase inhibitors in cultured human breast cancer cells

The presence of aromatase activity, estrogen receptors, and estrogenic responsiveness in MCF-7 human breast cancer cells has allowed this cell line to be used as a unique in vitro system for investigating the biological activities of potentially therapeutic aromatase inhibitors. We now report the results of studies which have examined the cytotoxicity, antiaromatase, and intrinsic estrogenic activities of aminoglutethimide, 1,2-dehydrotestolactose (testolactone), dihydrotestosterone, 4-hydroxy-4-androstene-3,17-dione, and 10-propargylestr-4-ene-3,17-dione r271w4.r272w1 =[17α-methyl-³H]-17α,21-dimethyl-19-norpregna-4,9-diene- 3,20-dione r277w1.r278w2 =(10-propargylestr-4-ene-3,17-dione>4-hydroxy-4-androstene- 3,17-dione>>dihydrotestosterone>>testolactone with MCF-7 monolayer cultures. Cell viability was determined by trypan blue exclusion, and aromatase activity was assessed by quantifying the amounts of [³H]estradiol formed from [³H]testosterone. Estrogenic activity was assessed by examining the ability of each inhibitor to increase cytoplasmic progesterone receptor and deplete cytoplasmic estrogen receptor concentrations in these cells during a 5-day incubation period. Cytoplasmic progesterone and estrogen receptors were measured by the single-saturating-dose technique using [17α-methyl-³H]-17α, 21-dimethyl-19-norpregna-4,9-diene-3,20-dione and [³H]estradiol as the labeled ligands for each assay, respectively. The results showed that all of these compounds were noncytotoxic aromatase inhibitors in MCF-7 cells but that these agents demonstrated marked differences in inhibitory potency (10-propargylestr-4-ene-3,17-dione>4-hydroxy-4androstene-3,17-dione>>dihydrote stosterone>>testolactone = aminoglutethimide). The incubation of cells with 4-hydroxy-4-androstene-3,17-dione resulted in cytoplasmic progesterone and estrogen receptor responses that were similar in magnitude to those observed in other cultures incubated with equimolar concentrations of estradiol. None of the other four agents demonstrated estrogenic activity in this system. However, we have previously observed that dihydrotestosterone has substantial antiestrogenic action in this system. Taken together, these results indicate that some aromatase inhibitors may influence the hormonal regulation of human breast cancer cells by more than one mechanism.