The hydrolysis of estrone sulfate and dehydroepiandrosterone sulfate by MCF-7 human breast cancer cells.
Reports of estrone (E1) and dehydroepiandrosterone (DHEA) sulfatase (sulfohydrolase) activities within many human breast cancers have prompted us to undertake the identification and partial characterization of these enzyme activities within MCF-7 human breast cancer cells. Enzyme assays were performed within subcellular preparations and intact cultures by quantifying the total nonpolar 3H-labeled metabolites formed from [3H]E1 sulfate (E1S) and [3H]DHEA sulfate (DHEAS). The results have shown that the hydrolysis of each steroid sulfate is mediated by different particulate enzymes, which demonstrate optimal activity between pH 6.0-7.0. The analysis of enzyme kinetic data showed the Km values of E1S and DHEAS for their enzymes to be approximately 6.3 and 3.6 microM/L, respectively. Neither enzyme was subject to product inhibition. Androsterone sulfate and pregnenolone sulfate produced significant inhibition of E1, but not DHEA, sulfatase activity. E1S inhibited DHEA sulfatase competitively, with an approximate Ki of 11 microM, whereas DHEAS inhibited E2 sulfatase in a noncompetitive fashion, demonstrating an approximate Ki of 0.6 microM. Studies carried out with intact MCF-7 cultures using physiological concentrations of 3H-labeled E1S (2 nM) or DHEAS (1 microM) showed the accumulation of nonpolar metabolites during a 20-h incubation period. When cultures were incubated with similar concentrations of both steroid sulfates the apparent intracellular activity of E1 sulfatase was reduced by approximately 70%, whereas DHEA sulfatase activity remained unchanged. The results of these studies confirm the ability of MCF-7 cells to hydrolyze extracellular E1S and DHEAS, indicate that these reactions are mediated by different enzymes, and demonstrate that DHEAS is a potent inhibitor of MCF-7 E1 sulfatase. Circulating DHEAS, therefore, may substantially limit the ability of most postmenopausal breast cancers to use E1S as a substrate for intracellular estrogen biosynthesis.
MacIndoe
J H
JH
Department of Internal Medicine, University of Iowa College of Medicine, Iowa City.
D001943:Breast Neoplasms / Q000378:metabolism*; D003687:Dehydroepiandrosterone / Q000031:analogs & derivatives* / Q000378:metabolism; D019314:Dehydroepiandrosterone Sulfate; D004970:Estrone / Q000031:analogs & derivatives* / Q000378:metabolism; D005260:Female; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D043266:Steryl-Sulfatase; D013429:Sulfatases / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000378:metabolism
D004970:Estrone; D003687:Dehydroepiandrosterone; D019314:Dehydroepiandrosterone Sulfate; D013429:Sulfatases; D043266:Steryl-Sulfatase; C017296:estrone sulfate