Modulation of steroidogenic gene expression and hormone production of H295R cells by pharmaceuticals and other environmentally active compounds.
The H295R cell bioassay was used to evaluate the potential endocrine disrupting effects of 18 of the most commonly used pharmaceuticals in the United States. Exposures for 48 h with single pharmaceuticals and binary mixtures were conducted; the expression of five steroidogenic genes, 3betaHSD2, CYP11beta1, CYP11beta2, CYP17 and CYP19, was quantified by Q-RT-PCR. Production of the steroid hormones estradiol (E2), testosterone (T) and progesterone (P) was also evaluated. Antibiotics were shown to modulate gene expression and hormone production. Amoxicillin up-regulated the expression of CYP11beta2 and CYP19 by more than 2-fold and induced estradiol production up to almost 3-fold. Erythromycin significantly increased CYP11beta2 expression and the production of P and E2 by 3.5- and 2.4-fold, respectively, while production of T was significantly decreased. The beta-blocker salbutamol caused the greatest induction of CYP17, more than 13-fold, and significantly decreased E2 production. The binary mixture of cyproterone and salbutamol significantly down-regulated expression of CYP19, while a mixture of ethynylestradiol and trenbolone, increased E2 production 3.7-fold. Estradiol production was significantly affected by changes in concentrations of trenbolone, cyproterone, and ethynylestradiol. Exposures with individual pharmaceuticals showed the possible secondary effects that drugs may exert on steroid production. Results from binary mixture exposures suggested the possible type of interactions that may occur between drugs and the joint effects product of such interactions. Dose-response results indicated that although two chemicals may share a common mechanism of action the concentration effects observed may be significantly different.
Gracia
Tannia
T
Department of Zoology, National Food Safety and Toxicology Center, Center for Integrative Toxicology, Michigan State University, East Lansing, MI 48824, USA. tg3@sanger.ac.uk
Hilscherova
Klara
K
Jones
Paul D
PD
Newsted
John L
JL
Higley
Eric B
EB
Zhang
Xiaowei
X
Hecker
Markus
M
Murphy
Margaret B
MB
Yu
Richard M K
RM
Lam
Paul K S
PK
Wu
Rudolf S S
RS
Giesy
John P
JP
D015096:3-Hydroxysteroid Dehydrogenases / Q000187:drug effects / Q000235:genetics; D018268:Adrenocortical Carcinoma / Q000378:metabolism*; D001141:Aromatase / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D019405:Cytochrome P-450 CYP11B2 / Q000187:drug effects / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000187:drug effects* / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D052244:Endocrine Disruptors / Q000494:pharmacology*; D004958:Estradiol / Q000096:biosynthesis; D005786:Gene Expression Regulation / Q000187:drug effects*; D012739:Gonadal Steroid Hormones / Q000096:biosynthesis*; D006801:Humans; D011374:Progesterone / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013252:Steroid 11-beta-Hydroxylase / Q000187:drug effects / Q000235:genetics; D013254:Steroid 17-alpha-Hydroxylase / Q000187:drug effects / Q000235:genetics; D013739:Testosterone / Q000096:biosynthesis