Novotnik Breda, Ščančar Janez, Milačič Radmila, Filipič Metka, Žegura Bojana
Department of Environmental Sciences, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia; Jožef Stefan International Postgraduate School, Jamova 39, 1000 Ljubljana, Slovenia.
National Institute of Biology, Department for Genetic Toxicology and Cancer Biology, Večna pot 111, Ljubljana, Slovenia.
Chemosphere. 2016 Jul;154:124-131. doi: 10.1016/j.chemosphere.2016.03.118. Epub 2016 Apr 1.
Chromium (Cr) and ethylenediaminetetraacetate (EDTA) are common environmental pollutants and can be present in high concentrations in surface waters at the same time. Therefore, chelation of Cr with EDTA can occur and thereby stable Cr(III)-EDTA complex is formed. Since there are no literature data on Cr(III)-EDTA toxicity, the aim of our work was to evaluate and compare Cr(III)-EDTA cytotoxic and genotoxic activity with those of Cr(VI) and Cr(III)-nitrate in human hepatoma (HepG2) cell line. First the effect of Cr(VI), Cr(III)-nitrate and Cr(III)-EDTA on cell viability was studied in the concentration range from 0.04 μg mL(-1) to 25 μg mL(-1) after 24 h exposure. Further the influence of non-cytotoxic concentrations of Cr(VI), Cr(III)-nitrate and Cr(III)-EDTA on DNA damage and genomic stability was determined with the comet assay and cytokinesis block micronucleus cytome assay, respectively. Cell viability was decreased only by Cr(VI) at concentrations above 1.0 μg mL(-1). Cr(VI) at ≥0.2 μg mL(-1) and Cr(III) at ≥1.0 μg mL(-1) induced DNA damage, while after Cr(III)-EDTA exposure no formation DNA strand breaks was determined. Statistically significant formation of micronuclei was induced only by Cr(VI) at ≥0.2 μg mL(-1), while no influence on the frequency of nuclear buds nor nucleoplasmic bridges was observed at any exposure. This study provides the first evidence that Cr(III)-EDTA did not induce DNA damage and had no influence on the genomic stability of HepG2 cells.
铬(Cr)和乙二胺四乙酸(EDTA)是常见的环境污染物,可能同时在地表水中以高浓度存在。因此,Cr与EDTA之间会发生螯合作用,从而形成稳定的Cr(III)-EDTA络合物。由于目前尚无关于Cr(III)-EDTA毒性的文献数据,我们研究的目的是评估并比较Cr(III)-EDTA与人肝癌(HepG2)细胞系中Cr(VI)和Cr(III)-硝酸盐的细胞毒性和遗传毒性活性。首先,在暴露24小时后,研究了Cr(VI)、Cr(III)-硝酸盐和Cr(III)-EDTA在0.04 μg mL(-1)至25 μg mL(-1)浓度范围内对细胞活力的影响。进一步分别用彗星试验和胞质分裂阻滞微核细胞试验测定了非细胞毒性浓度的Cr(VI)、Cr(III)-硝酸盐和Cr(III)-EDTA对DNA损伤和基因组稳定性的影响。仅在浓度高于1.0 μg mL(-1)时,Cr(VI)才会降低细胞活力。≥0.2 μg mL(-1)的Cr(VI)和≥1.0 μg mL(-1)的Cr(III)会诱导DNA损伤,而在Cr(III)-EDTA暴露后未检测到DNA链断裂的形成。仅在≥0.2 μg mL(-1)的Cr(VI)诱导下,微核形成具有统计学意义,而在任何暴露条件下均未观察到对核芽频率和核质桥的影响。本研究首次提供证据表明,Cr(III)-EDTA不会诱导DNA损伤,且对HepG2细胞的基因组稳定性没有影响。