PLHC-1세포주의 Comet assay를 이용한 하천 퇴적토의 생태독성평가
Studies on Toxicological Evaluation of Freshwater Sediment using a PLHC-1 Cell Comet Assay
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초록
본 연구에서는 환경오염 진단을 목적으로 어류간암세포에서 유래된 PLHC-1 세포를 이용하여 하천 퇴적토를대상으로 한 Comet assay (DNA 손상 평가기법)를 시도하였으며, NR assay 및 EROD induction과 병행함으로써,퇴적토의 생태독성 수준을 다양하게 평가 할 수 있는PLHC-1 세포시험기법을 확립하고자 하였다. 도시하천으로부터 채취된 7개 정점 하상퇴적토의 독성을 평가하였으며 생존율에서는 모든 정점에서 영향이관찰되지 않았다. 반면, EROD 역가와 Comet assay 평가에서는 공단이 인접한 정점들의 독성수준이 높게 나타났으며 정점별 특이성을 관측 할 수 있었다. 또한, PLHC-1세포와 올챙이 혈구세포의 Comet assay를 시행한 결과,퇴적토 유기용매 추출물 시료에 대하여 PLHC-1 세포의민감도가 높게 나타났다. 이러한 결과들은 기존의 세포 생존율과 올챙이 혈구세포의 Comet assay에 비교하여 PLHC-1 세포의 Comet assay와 EROD 역가 측정이 하천퇴적토 생태독성평가를비롯한 환경오염진단 과정에 평가종말점(Endpoint)으로서 활용이 적합한 것으로 판단되며, 독성이 높게 나타난문제 정점에 대하여 보다 집중적인 연구가 필요하다 사료된다.
Abstract
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.
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