Evidence for DNA-diquat interaction and cytotoxicity in in vitro rat cells |
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Authors: | Quan Zhang Cui Wang Wanpeng Liu Xiaofeng Zhang Shulin Zhuang |
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Institution: | (1) College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou, 310032, People’s Republic of China;(2) Institute of Environmental Science, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou, 310058, People’s Republic of China; |
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Abstract: | Over 98% of sprayed insecticides and 95% of herbicides reach non-target species in air, soil, and water. Numerous studies
have reported that pesticide residues can cause acute and chronic toxicity. Pesticide residues can be carcinogenic, mutagenic,
and immunotoxic. There are actually too few studies that bridge the disciplines of chemobioanalysis and environmental toxicology.
Here, we assessed the cytotoxicity of a bipyridilium herbicide diquat in rat adrenal pheochromocytoma cells (PC12). Our results
show that diquat caused the decrease in cell viability with a lethal concentration 50 (LC50) of 1.4 × 10−5 mol/L. This cytotoxicity may result from diquat-induced apoptosis, characterized by nuclear fragmentation and chromatin condensation
by Hoechst 33324 staining. To explore the possible mechanisms, the interaction between herbicide diquat and calf thymus DNA
(ctDNA) was further investigated using fluorescence quenching. The detection of static quenching showed that diquat was linked
with ctDNA by electrostatic interaction with a binding constant of 9.288 × 104 L/mol. This is the first study on the interaction of DNA with herbicide diquat by fluorometric method as well as on the evaluation
of cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Hoechst staining. Given the widespread
use of synthetic pesticides, the data would be valuable for the risk assessment of pesticide residues. |
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