Simple approach to measure metabolic pathways of steroids in living cells

Androgen metabolism and biotransformation in nontumoral and malignant human liver tissues and cells

“…The methods and procedures used to assess metabolic rates and pathways of sex steroids (intact cell analysis) have been originally established and optimized in our laboratories [16][17][18]. Briefly, cell or tissue cultures were washed twice with phosphate-buffered saline (PBS)-A and incubated for either 24 or 72 h in FCS-free, phenol red-free RPMI medium containing 1 nM tritiated androgen ([1,2,6,7-3 H(N)]-testosterone, S.A. 92.4 Ci/mmol, or [1,2,6,7-3 H(N)]-androstenedione, S.A. 84.5 Ci/mmol; DuPont de Nemours Italiana SpA, Milan, Italy) as precursor.…”

Sex Steroids, Carcinogenesis, and Cancer Progression

“…Aiming to assess rates and pathways of sex steroid metabolism, either in vivo or in vitro , we originally developed, established, and optimized a novel approach we named intact cell analysis 4,16. In this method, the use of reverse‐phase high performance liquid chromatography (RP‐HPLC) with online radioactive detection allows a simultaneous and sequential estimation of several enzyme activities in intact cultured cells or tissue fragments incubated for variable time lengths (from a few minutes up to several days) with physiological amounts of a radioactive (tritiated) sex steroid used as precursor.…”

“…Subsequently, we showed that unusual metabolites are excreted in markedly greater amounts in endometrial cancer patients whose tumors are estrogen receptor (ER)‐negative 3. Using an original intact cell analysis,4 we investigated sex steroid metabolism in a panel of human cancer cell lines, either hormone responsive or unresponsive, originating from human breast, endometrium, and prostate. We found that highly divergent patterns of steroid metabolism exist and that the catalytic preference (predominantly reductive or oxidative) is strictly associated with the steroid receptor status of cells 5–7.…”

Pharmacokinetics of Exogenous Natural and Synthetic Estrogens and Antiestrogens

“…Other methods described include a chemiluminescent immunoassay (SATO et al 1996); receptor-binding assays (IIDA et al 1991); high-performance liquid chromatography with photometric detection (CASTAGNETTA et al 1991;DESTA 1988;SUKKER et al 1981;TOWNSEND et al 1988); chemiluminescence detection (NOZAKI et al 1988); electrochemical detection (NOMA et al 1991) or ftuorimetric detection (KATAYAMA and TANIGUCHI 1993;CHANDRASEKARAN et al 1996); adsorptive stripping voltammetry (Hu et al 1992); recombinant cell bioassay (KLEIN et al 1994); and gas chromatography-mass spectrometry (GC-MS) (SPINK et al 1990;VANLUCHENE et a1.1983;ZIMMERMANN et al 1998b).…”