Chinese Hamster Ovary 세포에서 2-aminoanthracene의 유전독성 측정을 위한 Single Cell Gel Electrophoresis 조건
Conditions of Single Cell Gel Electrophoresis of Chinese Hamster Ovary Cell for Detect the Genetic Toxicity of 2-aminoanthracene
박장환, 김동원,
소속 상세정보
박장환 ( Park Chang-Hwan )
한양대학교 의과대학 미생물학교실
김동원 ( Kim Dong-Won )
한양대학교 의과대학 마취통증의학교실
KMID : 0603520010060020092
Abstract
The single cell gel electrophoresis (SCGE, comet assay) is becoming established as a genotoxicity test with manifold applications in vitro and in vivo. While the underlying principles are identical, various modifications of the method are in use which clearly affect the sensitivity and resolving power of the assay. We therefore performed SCGE experiments with CHO.1A2BRN2E cells, which express human cytochrome p450 1A2 cytochrome p450 reductase and N-acetyltransferase 2 genes, using different unwinding time, voltage/ampere, electrophoresis time and pH of electrophoresis buffer. DNA damage was induced by 2-aminoanthracene (2-AA) during 24 hours. On the basis of results obtained in this study, best SCGE conditions for detect DNA damage induced 2-AA to CHO.1A2BRN2E cells are 30 minutes unwindings, 25 volt/ 300 ㎃, 20 minutes electrophoresis with alkali electrophoresis buffer (pH 13.5).
키워드
Single cell gel electrophoresis (SCGE); CHO cell; Metabolic activation
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