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Both pentachlorophenol and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) had been studied widely because of their probable anti-estrogenic activity. Sodium pentachlorophenol (PCP-Na), as a industrial product used in many fields, usually contains a trace of TCDD. The aim of this study was to assess the anti-estrogenic effect of PCP-Na in juvenile goldfish (Carassius auratus)hepatocyte cultures using vitellogenin (VTG) as the biomarker. The ID50 of PCP-Na was investigated and then a series of concentrations (0.001-0.5 μg/ml) of PCP-Na were evaluated to estimate the anti-estrogenic activity. Results showed that PCP-Na was cytotoxic for hepatocytes even at very low concentration ＜1.21 μg/ml, and it could not induce VTG at any concentrations tested.Since it failed to stimulate VTG production, the possibility of its anti-estrogenic effect was tested, and a well-known anti-estrogenic compound-tamoxifen was used as positive control. PCP-Na caused a reduction in VTG synthesis in juvenile goldfish (Carassius auratus) hepatocytes at concentrations ＞0.1 μg/ml when co-exposure with 1 μg/ml 17β-estradiol (E2), making its anti-estrogenic activity approximately as potent as tamoxifen. Our results indicate that PCP-Na can act as negative modulators of estrogenic function in juvenile goldfish (Carassius auratus) hepatocytes.