title: An in vitro high-throughput assay for screening reproductive and toxic effects of anticancer compounds.
abstract: An in vitro assay was developed that simultaneously tested the effects of anticancer drug candidates on cytotoxicity, hormone synthesis, and gonadotrophin responsiveness using the choriocarcinoma JAr cell line. JAr culture conditions were optimized and then cells were exposed to a marine mollusc extract in the presence and absence of hCG. The intra- and interassay coefficients of variation of the optimized 1 H thiazolyl blue tetrazolium bromide assay were 11.3% and 10.9%, respectively. hCG (1,000 mIU/mL) increased progesterone (P4) synthesis after 24 H (P<0.05). The mollusc extract significantly decreased cell viability, with the IC50 affected by incubation time, but not hCG. P4 synthesis was inhibited at low concentrations of the anticancer extract, but stimulated at the highest concentration, and complex interactions of P4 were also found with hCG. In conclusion, the optimized assay is useful to characterize the effects of novel drugs on cytotoxicity, basal, and gonadotrophin-stimulated P4 synthesis in vitro, and can be used to inform subsequent in vivo studies.
authors.affiliation: Department of Medical Biotechnology, School of Medicine, Flinders University Adelaide, South Australia, Australia.
authors.forename: Vicki
authors.initials: V
authors.lastname: Edwards
authors.forename: Kirsten
authors.initials: K
authors.lastname: Benkendorff
authors.forename: Fiona
authors.initials: F
authors.lastname: Young
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology / Q000633:toxicity; D001688:Biological Products / Q000494:pharmacology / Q000633:toxicity; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects*; D006063:Chorionic Gonadotropin / Q000032:analysis* / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology / Q000633:toxicity; D008974:Mollusca / Q000737:chemistry; D011374:Progesterone / Q000032:analysis* / Q000378:metabolism; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D000970:Antineoplastic Agents; D001688:Biological Products; D006063:Chorionic Gonadotropin; D011374:Progesterone; D006861:Hydrogen Peroxide
keywords: JAr cells; brominated indoles; cytotoxicity assay; human choronic gonadotrophin; marine natural products; progesterone