氯化饮水有机提取物诱导人胚肝细胞(L-02)DNA损伤与RAS基因表达的研究  

Study on DNA damage and expression of RAS gene in L-02 cells exposed to chlorinated drinking water extracts

作　　者：刘慧[1] 邹亚玲[1] 刘爱林[1] 李晓燕[1] 鲁文清[1] 

机构地区：[1]华中科技大学同济医学院公共卫生学院,武汉430030

出　　处：《卫生研究》2004年第5期531-533,共3页Journal of Hygiene Research

基　　金：国家自然科学基金项目 (No .30 1 70 794)

摘　　要：目的　研究武汉市主要水源D湖氯化饮水有机提取物对人胚肝细胞 (L 0 2 )DNA损伤与RAS基因表达的诱导 ,探讨氯化饮水有机提取物诱导遗传损伤的分子机制。方法　以L 0 2细胞作为靶细胞 ,应用单细胞凝胶电泳试验 (SCGE)测定DNA链断裂及用原位杂交试验 (ISH)测定RAS基因表达。氯化饮水有机提取物设 0 16 7、1 6 7、16 7、16 7水样ml ml培养液 4个剂量 ,DMSO(10ml L)为溶剂对照 ,B(a)P(2 0 0 μmol L)为阳性对照。结果　与溶剂对照相比 ,氯化饮水有机提取物在较高剂量组 (16 7和 16 7L L水样培养液 )能引起DNA链断裂程度的明显增加 ;不同浓度的氯化饮水有机提取物均可使RAS基因表达水平明显增加。氯化饮水有机提取物在 0 16 7、1 6 7、16 7L L水样培养液的浓度范围内 ,其RAS基因表达水平和DNA迁移度呈高度正相关 (r=0 99)。结论　氯化饮水有机提取物可诱导L 0 2细胞DNA损伤与RAS基因表达 。Objective To study the DNA damage and expression of RAS gene in human fetal hepatocytes (L-02) induced by chlorinated drinking water extracts, and to explore the possible molecular mechanism of its genotoxicity .Methods L-02 cells were exposed to chlorinated drinking water extracts at the concentration of 0.167,1.67,16.7 and 167ml /ml culture medium. Single cell gel electrophoresis (SCGE) and in situ hybridization (ISH) assay were used to detect DNA damage and the expression of RAS gene in L-02 cells. DMSO(10ml/L)and B(a)P(200μmol/L)were used as solvent control and positive control respectively. Results Chlorinated drinking water extracts led to a significant increase of DNA migration in the L-02 cells at the concentrations of 16.7 and 167ml/ml culture medium in comparison with solvent control. Significantly higher expressions of RAS gene was found in the L-02 cells exposed to all concentrations of chlorinated drinking water extracts than in that of solvent control group. There was a significant correlation (r=0.99) between RAS gene expression and DNA migration when the concentration of chlorinated drinking water extracts was between 0.167 and 16.7ml/ml. Conclusion Chlorinated drinking water extracts could induce obvious DNA damage and RAS gene expression in L-02 cells, and RAS gene expression might be affected by DNA damage.

关 键 词：氯化饮水有机提取物 人胚肝细胞 单细胞凝胶电泳试验 原位杂交试验 DNA损伤 RAS基因 

分 类 号：R123.6[医药卫生—环境卫生学] R730.231[医药卫生—公共卫生与预防医学]