Jun 2, 2020

Cytotoxic and genotoxic evaluation of cotinine using human neuroblastoma cells (SH-SY5Y)

Genetics and Molecular Biology
Daiana DalbertoJuliana da Silva

Abstract

Cotinine is the main metabolite of nicotine, which is metabolized in the liver through a cytochrome P450 enzyme. Different studies point to genetic instability caused by nicotine, such as single and double DNA strand breaks and micronuclei formation, but little is known about the effect of cotinine. Therefore, the present in vitro study assessed the effects of cotinine on cell viability and DNA damage in SH-SY5Y neuroblastoma cells, as well as genotoxicity related to oxidative stress mechanisms. Comparisons with nicotine were also performed. An alkaline comet assay modified by repair endonucleases (FPG, OGG1, and Endo III) was used to detect oxidized nucleobases. SH-SY5Y neuronal cells were cultured under standard conditions and exposed for 3 h to different concentrations of cotinine and nicotine. Cytotoxicity was observed at higher doses of cotinine and nicotine in the MTT assay. In the trypan blue assay, cells showed viability above 80% for both compounds. Alkaline comet assay results demonstrated a significant increase in damage index and frequency for cells treated with cotinine and nicotine, presenting genotoxicity. The results of the enzyme-modified comet assay suggest a DNA oxidative damage induced by nicotine. Unlike ot...Continue Reading

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Mentioned in this Paper

Oxidized DNA Binding
Structure
Trypan Blue Stain Method
Mutagenicity Tests
Cytotoxicity
Metabolite
Study
Cotinine
DNA Damage
Micronuclei, Chromosome-Defective

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