Oil sands process-affected water (OSPW) have been reported to exhibit endocrine disrupting effects on aquatic organisms. Although the responsible compounds are unknown, naphthenic acids (NAs) have been considered to be implicated. The current study was designed to investigate the endocrine disruption of OSPW extracted NAs (OS-NAs) and commercial NAs (C-NAs) using a combination of in vitro and in vivo assays. The effects of OS-NAs and C-NAs on steroidogenesis were assessed both at hormone levels and expression levels of hormone-related genes in the H295R cells. The transcriptions of biomarker genes involved in endocrine systems in zebrafish larvae were investigated to detect the effects of OS-NAs and C-NAs on endocrine function in vivo. Exposure to OS-NAs and C-NAs significantly increased production of 17 beta-estradiol (E2) and progesterone (P4), and decreased production of testosterone (T). Both OS-NAs and C-NAs significantly induced the expression of several genes involved in steroidogenesis. The abundances of transcripts of biomarker gene CYP19b, ER alpha, and VTG were significantly up-regulated in zebrafish larvae exposed to OS-NAs and C-NAs, which indicated that NAs had negative effects on estrogen-responsive gene transcription in vivo. These results indicated that NAs should be partly responsible for the endocrine disrupting effects of OSPW. (C) 2015 Elsevier B.V. All rights reserved.