Environmental Monitoring and Assessment

, Volume 175, Issue 1–4, pp 315–319 | Cite as

A flounder (Paralichthys olivaceus) gill cell line as in vitro acute assay system of nonylphenol cytotoxicity



A cell line (FG cells) derived from a gill of the flounder, Paralichthys olivaceus were used to determine the cytotoxic effects of nonylphenol (NP). Cytotoxicity was measured by three endpoint systems: neutral red (NR) uptake assay, methyl thiazolyl tetrazolium (MTT) assay and cell protein assay. The result showed that NP was cytotoxic to FG cells at all tested concentrations, and toxicity increased as the concentration of NP was progressively increased. The 24 h-IC50 values of NP were 39.81, 37.76 and 38.22 μmol/L for NR uptake, MTT assay and cell protein assay, respectively. Moreover, the morphological changes of FG cells were also studied at the concentration of 30 μmol/L for 24 h. Cells morphology were markedly altered by NP observed under a scanning electron microscopy, as evidenced by swelling cells, two and more nucleolus and an increased number of lipid particles. This would suggest that the FG cell line is a suitable bioindicator for the screening of the acute toxicity of NP.


Cytotoxicity FG cells Nonylphenol Paralichthys olivaceus 


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  1. Aoki, M., Kurasaki, M., Saito, T., Seki, S., Hosokawa, T., Takahashi, Y., et al. (2004). Nonylphenol enhances apoptosis induced by serum deprivation in PC12 cells. Life Sciences, 74(18), 2301–2312.CrossRefGoogle Scholar
  2. Arukwe, A., Thibaut, R., Ingebrigtsen, K., Celius, T., Goksøyr, A., & Cravedi, J.-P. (2000). In vivo and in vitro metabolism and organ distribution of nonylphenol in Atlantic salmon (Salmo salar). Aquatic Toxicology, 49(4), 289–304.CrossRefGoogle Scholar
  3. Babich, H., & Borenfreund, E. (1987). In vitro cytotoxicity of organic pollutants to bluegill sunfish (BF-2) cells. Environmental Research, 42(1), 229–237.CrossRefGoogle Scholar
  4. Blackburn, M. A., & Waldock, M. J. (1995). Concentrations of alkylphenols in rivers and estuaries in England and Wales. Water Research, 29(7), 1623–1629.CrossRefGoogle Scholar
  5. Bols, N. C., Boliska, S. A., & Dixon, D. G. (1985). Use of fish cell cultures as an indication of contaminant toxicity to fish. Aquatic Toxicology, 6(2), 147–155.CrossRefGoogle Scholar
  6. Borenfreund, E., & Puerner, J. A. (1985). Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicology Letters, 24(2–3), 119–124.CrossRefGoogle Scholar
  7. Borenfreund, E., Babich, H., & Martin-Alguacil, N. (1988). Comparison of two in vitro cytotoxicity assays: The neutral red (NR) and tetrozolium (MTT) tests. Toxicology in Vitro, 2(1), 1–6.CrossRefGoogle Scholar
  8. Ekwall, B. (1995). The basal cytotoxicity concept. In A. Goldberg & L. Zutphen (Eds.), Alternative methods in toxicology (pp, 721–725). New York: Mary Ann Liebert.Google Scholar
  9. Hale, R. C., Smith, C. L., De Fur, P. O., Harvey, E., Bush, E. O., La Guardia, M. J., et al. (2000). Nonylphenols in sediments and effluents associated with diverse wastewater outfalls. Environmental Toxicology and Chemistry, 19(4), 946–952.CrossRefGoogle Scholar
  10. Hughes, P. J., McLellan, H., Lowes, D. A., Zafar Kahn, S., Bilmen, J. G., Tovey, S. C., et al. (2000). Estrogenic alkylphenols induce cell death by inhibiting testis endoplasmic reticulum Ca2 +  pumps. Biochemical and Biophysical Research Communications, 277(3), 568–574.CrossRefGoogle Scholar
  11. Huuskonen, S. E., Hahn, M. E., & Lindström-Seppä, P. (1998). A fish hepatoma cell line (PLHC-1) as a tool to study cytotoxicity and CYP1A induction properties of cellulose and wood chip extracts. Chemosphere, 36(14), 2921–2932.CrossRefGoogle Scholar
  12. Kinnberg, K., Korsgaard, B., & Bjerregaard, P. (2000). Concentration-dependent effects of nonylphenol on testis structure in adult platyfish Xiphophorus maculatus. Marine Environmental Research, 50(1–5), 169–173.CrossRefGoogle Scholar
  13. Kobayashi, K., Tamotsu, S., Yasuda, K., & Oishi, T. (2005). Vitellogenin-immunohistochemistry in the liver and the testis of the Medaka, Oryzias latipes, exposed to 17β-estradiol and p-nonylphenol. Zoological Science, 22(4), 453–461.CrossRefGoogle Scholar
  14. Li, H. Y., & Zhang, S. C. (2001). In vitro cytotoxicity of the organophosphorus pesticide parathion to FG-9307 cells. Toxicology in Vitro, 15(6), 643–647.CrossRefGoogle Scholar
  15. Marion, M., & Denizeau, F. (1983). Rainbow trout and human cells in culture for the evaluation of the toxicity of aquatic pollutants. A study with lead. Aquatic Toxicology, 3(1), 47–60.CrossRefGoogle Scholar
  16. Na, N., Guo, H. R., Zhang, S. C., Li, Z. J., & Yin, L. C. (2009). In vitro and in vivo acute toxicity of fenpyroximate to flounder Paralichthys olivaceus and its gill cell line FG. Aquatic Toxicology, 92(2), 76–85.CrossRefGoogle Scholar
  17. Rachlin, J. W., & Perlmutter, A. (1968). Fish cells in culture for study of aquatic toxicants. Water Research, 2(6), 409–414.CrossRefGoogle Scholar
  18. Saito, H., Iwami, S., & Shigeoka, T. (1991). In vitro cytotoxicity of 45 pesticides to goldfish GF-scale (GFS) cells. Chemosphere, 23(4), 525–537.CrossRefGoogle Scholar
  19. Shopsis, C., & Eng, B. (1985). Rapid cytotoxicity testing using a semiautomated protein determination on cultured cells. Toxicology Letters, 26(1), 1–8.CrossRefGoogle Scholar
  20. Solé, M., López de Alda, M. J., Castillo, M., Porte, C., Ladegaard-Pedersen, K., & Barceló, D. (2000). Estrogenicity determination in sewage treatment plants and surface waters from the Catalonian area (NE Spain). Environmental Science & Technology, 34(24), 5076–5083.CrossRefGoogle Scholar
  21. Tong, S. L., Li, H., & Miao, H. Z. (1997). The establishment and partial characterization of a continuous fish cell line FG-9307 from the gill of flounder Paralichthys olivaceus. Aquaculture, 156, 327–333.CrossRefGoogle Scholar
  22. Wang, X., Han, X., Hou, Y., Yao, G., & Wang, Y. (2003). Effect of nonylphenol on apoptosis of Sertoli cells in vitro. Bulletin of Environmental Contamination and Toxicology, 70, 898–904.CrossRefGoogle Scholar
  23. Xiao, Q., Zhang, S. C., Hu, J. H., & Xu, Y. Y. (2006). Sperm of rosy barb (Puntius conchonius) as an in vitro assay system of nonylphenol cytotoxicity. Journal of Environmental Sciences, 18(3), 417–419.Google Scholar
  24. Xiao, Q., Zhang, S. C., Guo, H. R., Su, F., & Xu, Y. Y. (2007). Nonylphenol causes decrease in antioxidant enzyme activities, increase in O2 content and alteration in ultrastructures of FG cells, a flounder (Paralichthys olivaceus) gill cell line. Toxicology Mechanisms and Methods, 17(3), 127–134.CrossRefGoogle Scholar

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© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.College of OceanHebei Agricultural UniversityQinhuangdaoPeople’s Republic of China

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