Abstract
Acrylamide (AA) toxicity remains an interesting subject in toxicological research. The aim of the research performed in this paper was to determine mechanisms of cyto- and genotoxic effects of AA on the human colon adenocarcinoma cell line Caco-2, to estimate the inhibitory concentration (IC)50 values in cell viability assays, to measure the basal and oxidative DNA damage as well as the oxidative stress leading to apoptosis, and to assess the morphological changes in cells using microscopic methods. It has been proven that AA induces cytotoxic and genotoxic effects on Caco-2 cells. Higher cytotoxic activity was gained in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay compared with the PrestoBlue assay, with IC50 values of 5.9 and 8.9 mM after 24 h exposure, respectively. In the single-cell gel electrophoresis assay, the greatest DNA damage was caused by the highest concentration of acrylamide equal to 12.5 mM (89.1% ± 0.9%). AA also induced oxidative DNA damage and generated reactive oxygen species (ROS), which was concentration dependent and correlated with the depletion of mitochondrial membrane potential and apoptosis induction. In the microscopic staining of cells, AA in the dosage close to the IC50 induced morphological changes typical for apoptosis. Taken together, these results demonstrate that AA has a pro-oxidative effect on Caco-2 cells, leading to apoptotic cell death.
Highlights
It has been known for a long time that the formation of certain chemicals during food processing or preparation may pose a risk to human health
Treatment of Caco-2 cells with AA (0.2–50 mM) resulted in a time- and dose-dependent decrease in cell viability, as measured by both MTT and PrestoBlue assays after 24–72 h (Tables S1 and S2, Supplementary Materials)
Both assays showed that different concentrations of AA treatment significantly decreased Caco-2 cell proliferation at 24–72 h when compared with the AA unexposed negative control (p < 0.05)
Summary
It has been known for a long time that the formation of certain chemicals during food processing or preparation may pose a risk to human health. AA occurs in thermally processed food as a carcinogen—it is formed via the Maillard reaction when heating carbohydrate-rich foods at temperatures above 120 degrees. The presence of the compound in food is very substantial, owing to its mutagenic and carcinogenic properties. The International Agency for Research on Cancer (IARC) classified AA as a potentially carcinogenic substance for humans, as it possibly causes DNA damage and gene mutation [3]. It stated that AA is a probable human carcinogen, Molecules 2020, 25, 368; doi:10.3390/molecules25020368 www.mdpi.com/journal/molecules
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