title: Mimicking postmenopausal steroid metabolism in breast cancer cell culture: Differences in response to DHEA or other steroids as hormone sources.
abstract: Following menopause virtually 100% of estrogens are synthesized in peripheral target tissues from precursor steroids of adrenal origin. These steroids are the unique source of sex steroids in these women. This positions some steroid metabolizing enzymes as primary targets for novel therapies for estrogen receptor-positive (ER+) breast cancer. However, previous research on the steroid-converting enzymes has been performed using their direct substrate as a hormone source, depending on the facility where studied and the robust signal obtained. These experiments may not always provide an accurate reflection of physiological and post-menopausal conditions. We suggest providing dehydroepiandrosterone (DHEA) as an intracrinological hormone source, and comparing the role of steroid-converting enzymes using DHEA and their direct substrates when an extensive mechanistic understanding is required. Here, we present a comparative study of these enzymes with the provision of DHEA and the direct substrates, estrone (E1) or dihydrotestosterone (DHT), or additional steroids as hormone sources, in breast cancer cells. Enzyme knockdown by respective specific siRNAs and observations on the resulting differences in biological function were carried out. Cell biology studies showed no difference in biological function for 17β-HSD1 and 17β-HSD7 when cultured with different steroid hormones: cell proliferation and estradiol levels decreased, whereas DHT accumulated; cyclinD1, PCNA, and pS2 were down-regulated after knocking down these two enzymes, although the quantitative results varied. However, culture medium supplementation was found to have a marked impact on the study of 3α-HSD3. We demonstrated that provision of different steroids as a substrate or hormone sources may promote modified biological effects: provision of DHEA is the preferred choice to mimic postmenopausal steroid metabolism in cell culture.
authors.affiliation: Laboratory of Molecular Endocrinology and Oncology, Centre Hospitalier Universitaire de Québec Research Center (CHUQ-CHUL) and Department of Molecular Medicine, Laval University, 2705 boulevard Laurier, Québec G1V4G2, Canada.
authors.forename: Dan
authors.initials: D
authors.lastname: Xu
authors.affiliation: Laboratory of Molecular Endocrinology and Oncology, Centre Hospitalier Universitaire de Québec Research Center (CHUQ-CHUL) and Department of Molecular Medicine, Laval University, 2705 boulevard Laurier, Québec G1V4G2, Canada. Electronic address: sheng-xiang.lin@crchul.ulaval.ca.
authors.forename: Sheng-Xiang
authors.initials: SX
authors.lastname: Lin
mesh_terms: D015067:17-Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D017209:Apoptosis; D001940:Breast / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism*; D002453:Cell Cycle; D049109:Cell Proliferation; D003687:Dehydroepiandrosterone / Q000378:metabolism*; D013196:Dihydrotestosterone / Q000378:metabolism*; D004970:Estrone / Q000378:metabolism*; D005260:Female; D006801:Humans; D061986:MCF-7 Cells; D017698:Postmenopause / Q000378:metabolism*
chemical_list: D013196:Dihydrotestosterone; D004970:Estrone; D003687:Dehydroepiandrosterone; D015067:17-Hydroxysteroid Dehydrogenases; C044256:3 (or 17)-beta-hydroxysteroid dehydrogenase
keywords: 17β-HSD1; 17β-HSD7; 3α-HSD3; Breast cancer; DHEA; DHT; E2; siRNA knockdown