PLHC-1세포주의 Comet assay를 이용한 하천 퇴적토의 생태독성평가

Studies on Toxicological Evaluation of Freshwater Sediment using a PLHC-1 Cell Comet Assay

  • 박정아 (국립수산과학원 어장환경과) ;
  • 황인영 (인제대학교 환경학과) ;
  • 백승홍 (국립수산과학원 어장환경과) ;
  • 김영숙 (국립수산과학원 어장환경과)
  • Bak, Jeong-Ah (Marine Environment Research Division, National Fisheries Research and Development Institute) ;
  • Hwang, In-Young (Environmental Science & Engineering, Inje University) ;
  • Baek, Seung-Hong (Marine Environment Research Division, National Fisheries Research and Development Institute) ;
  • Kim, Young-Sug (Marine Environment Research Division, National Fisheries Research and Development Institute)
  • 투고 : 2010.10.26
  • 심사 : 2011.01.12
  • 발행 : 2011.02.28

초록

In this study, the Comet assay (evaluation of DNA damage) used the fish hepatocellular carinoma cell, PLHC-1, was tried to the sediment extract obtained from freshwater to understand its applicability as a tool for monitoring sediment toxicity. In parallel, induced EROD (7-ethoxyresorufin- O-deethylase) activity and DNA damage (TEM values) in PLHC-1 cells were measured for establishing the tandem endpoints of the PLHC-1cell test to test the ecotoxicity of sediment. Among several study sites in a small river passed through downtown and industrial park area, one of them, site B, showed a higher level of EROD activity and DNA damage than other sites. It indicates that a tandem endpoints of PLHC-1 cells could be useful tools for assessing the toxicity of sediment. The sensitivity of Comet assay with PLHC-1 cells was a little higher than that with a blood cell of frog tadpoles to the solvent extract of sediment. According to the results, a PLHC-1 cell-Comet assay could be used as a useful tool for evaluating ecotoxicity of the freshwater sediment. In addition, more detailed studies are needed to the contaminated site.

과제정보

연구 과제 주관 기관 : 국립수산과학원

참고문헌

  1. 고성룡, 정규혁. 2000. EROD-microbioassay에 의한 원천천 상류지역 수질 조사. 한국환경위생학회지. 26:105-109.
  2. 김기범, 배세진. 2003. Pyrene과 Benzo(a)pyrene에 노출된 굴의 혈구 아가미 세포에서의 DNA 손상 측정을 위한 Comet assay의 이용. 한국양식학회지. 16(3):196-201.
  3. 문성환, 조재구, 정진애, 백승홍, 안지영, 박미선, 박재호, 황인영. 2005. 낙동강 하류 유입수 수질의 환경독성학적 평가 및 비교. 한국환경독성학회 학술대회자료. pp. 88-90.
  4. 박선영, 최진희. 2006. Comet assay를 이용한 지표 생물종에서의 환경유해물질로 인한 유전 모니터링. 대한환경공학회 춘계학술연구발표회 논문집. pp. 1805-1810.
  5. 조홍제, 이병호, 이근배. 2000. 도시하천 하상퇴적토의 특성에 관한 연구. 대한토목학회논문집. pp. 607-618.
  6. 환경부. 2008. 공단주변 공공수역 생태위해성 평가체계 구축을 위한 시범사업.
  7. 환경부. 2009. 공단주변 공공수역 생태위해성 평가체계구축 및 위해원인 규명을 위한 연구사업.
  8. Babich H, JA Puerner and E Borenfreund. 1986. In vitro cytotoxicity of metals to bluegill (BF-2) cells. Archives of Environmental Contamination and Toxicology 15:31-37. https://doi.org/10.1007/BF01055246
  9. Borenfreund E and JA Puerner. 1985. Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicology Letters 24:119-124. https://doi.org/10.1016/0378-4274(85)90046-3
  10. Burke MD and RT Mayer. 1974. Ethoxyresorufin: direct fluorometric assay of a microsomal O-dealkylation which is preferentially inducible by 3-methylcholanthrene. Drug Metabolism, Disposition 2:583-588.
  11. Castano A, MJ Cantarino, P Castillo and JV Tarazona. 1996. Correlations between the RTG-2 cytotoxicity test $EC_{50}$ and in vivo $LC_{50}$ rainbow trout bioassay. Chemosphere 32: 2141-2157. https://doi.org/10.1016/0045-6535(96)00126-9
  12. Celander M, J Bremer and E Mark. 1997. Glucocorticoid-Xenobiotic interactions: Dexamethasone-Mediated potentiation of cytochrome P4501A induction by $\beta$-naphthoflavone in a fish hepatoma cell line (PLHC-1). Environmental Toxicology and Chemistry 16:900-907.
  13. Celander M, ME Hahn and JJ Stegeman. 1996. Cytochromes P450 (CYP) in the Poeciliopsis lucida hepatocellular carcinoma cell line (PLHC-1): dose- and time-dependent glucocorticoid potentiation of CYP1A induction without induction of CYP3A. Archives of Biochemistry and Biophysics 329: 113-122. https://doi.org/10.1006/abbi.1996.0198
  14. Chaparro RJF, U Kammann, H Steinhartb and N Theobald. 2000. Genotoxic potential of marine sediments from the North Sea. Mutation Research 467:161-168. https://doi.org/10.1016/S1383-5718(00)00030-9
  15. Fent K and J Hunn. 1996, Cytotoxicity of organic environmental chemicals to fish liver cells (PLHC-1). Marine Environmental Research 42:377-382. https://doi.org/10.1016/0141-1136(95)00043-7
  16. Gajski G, V Garaj-Vrhovac and V Orescanin. 2008. Cytogenetic status and oxidative DNA-damage induced by atorvastatin in human peripheral blood lymphocytes: Standard and Fpg-modified comet assay. Toxicology and Applied Pharmacology 231:85-93. https://doi.org/10.1016/j.taap.2008.03.013
  17. Haasch ML, EM Quardokus, LA Sutherland, MS Goodrich and JJ Lech. 1993. Hepatic CYP1A1 induction in rainbow trout by continuous flow thorough exposure to $\beta$-naphthoflavone. Fundamental Applied Toxicology 20:72-82. https://doi.org/10.1006/faat.1993.1009
  18. Hahn ME, BL Woodward, JJ Stegeman1 and SW Kennedy. 1996. Rapid assessment of induced cytochrome P4501A protein and catalytic in fish hepatoma cells grown in multiwell plates: response to TCDD, TCDF, and two planar PCBs. Environmental Toxicology and Chemistry 127:187-198.
  19. Huuskonen SE, TE Ristola, A Tuvikene, ME Hahn, JVK Kukkonen and P Lindstrom-Seppa. 1998. Comparison of two bioassays, a fish liver cell line (PLHC-1) and a midge (Chironomus riparius), in monitoring freshwater sediments. Aquatic Toxicology 44:47-67. https://doi.org/10.1016/S0166-445X(98)00070-8
  20. Jeong YN. 2007. Arsenic toxicity in PLHC-1 cell line and the distribution of arsenic in central appalachian. Marshall University.
  21. Jung DK, T Klaus and K Fent. 2001. Cytochrome P450 induction by nitrated polycyclic aroomatic hydrocarbons, azaarenes, and binary mixtures in fish hepatoma cell line PLHC- 1. Environmental Toxicology and Chemistry 20:149-159. https://doi.org/10.1002/etc.5620200117
  22. Kennedy SW, SP Jones and LJ Bastien. 1995. Efficient analysis of cytochrome P4501A catalytic activity, porphyrins and tatal proteins in chichken embryo hepatocytes cultures with a fluorescnece plate reader. Analytical Biochemistry 226: 362-370. https://doi.org/10.1006/abio.1995.1237
  23. Kwok YC, DPH Hsieh and PK Wong. 2005. Toxicity identification evaluation (TIE) of pore water of contaminated marine sediments collected from Hong Kong waters. Marine Pollution Bulletin 51:1085-1091. https://doi.org/10.1016/j.marpolbul.2005.06.009
  24. McCarthy JF and LR Shugart. 1990. Biomarkers of Environmental Contamination. Lewis Publisher. pp. 205-216.
  25. Pardos M, C Benninghoff, RL Thomas and S Khim-Heang. 1999. Confirmation of elemental sulfur toxicity in the Microtox${\circledR}$ assay during organic extracts assessment of freshwater sediments. Environmental Toxicology and Chemistry 18:188-193. https://doi.org/10.1002/etc.5620180213
  26. Phillips BM, BS Anderson, JW Hunt, SL Clark, JP Voorhees, RS Tjeerdema, J Casteline and M Stewart. 2009. Evaluation of phase II toxicity identification evaluation methods for freshwater whole sediment and interstitial water. Ecotoxicology and Environmental Safety. pp. 1-8.
  27. Putnam KP, DW Bombick and DJ Doolittle. 2002. Evaluation of eight in vitro assays for assessing the cytotoxicity of cigarette smoke condensate. Toxicology in Vitro 16:599-607. https://doi.org/10.1016/S0887-2333(02)00050-4
  28. Repetto G, A Jos, MJ Hazen, ML Molero, A del Peso, M Salguero, P del Castillo, MC Rodriguez-Vicente and M Repetto. 2001. A test battery for the ecotoxicological evaluation of pentachlorophenol. Toxicology in Vitro 15:503-509. https://doi.org/10.1016/S0887-2333(01)00055-8
  29. Saito H and T Shigeoka. 1994. Comparative cytotoxicity of chlorophenols to cultured fish cells. Environmental Toxicology and Chemistry 13:1649-1650. https://doi.org/10.1002/etc.5620131014
  30. Singh NP, MT McCoy, RR Tice and EL Schneider. 1988. A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research pp. 184-191.
  31. Stephen LW and LB Henry. 1969. The acute toxicity of some heavy metals to different species of aquatic insects. J. water. Poll. Ctrl. Fed. 41:280-284.