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Updated: Aug 29, 2026

A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage
Published on: May 10, 2022
[Effects of acrylamide on DNA damage in human keratinocytes]
Xiao-xia Ma1, Geng-dong Yao, Hao Cheng
1Institute of Occupational and Environmental Health, Zhejiang University, Hangzhou 310031, China.
Objective:
To investigate the toxic and DNA damaging effect of acrylamide (AA) on human keratinocytes and its mechanism.
Methods:
(1) After the keratinocyte cell line HaCaT cells were exposed to AA with different concentrations for 44 hours, cell survival rate was detected by MTT method. (2) The effects of DNA damage of exposed cells were detected by comet assay. (3) After treating the cells with 2.00 mmol/L of AA plus 0.50 mmol/L of 1-aminobenzotriazole (1-ABT), an inhibitor of cytochrome P-450 enzymes (CYP-450), for 4 hours, the relationship between DNA damage and CYP-450 was studied.
Results:
(1) Cytotoxicity measurement of AA showed that cell survival rate decreased significantly after 44-hour treatment. (2) Cytotoxicity was not detected after 4-hour AA treatment, but significant DNA damage was observed in all treatment groups, and the degree of damage increased with the concentration of AA. Moreover, the tail lengths of comet cells were in dose-effect relationship. As for cells treated by 1-ABT with 2 mmol/L AA, comet rate and tail length were 15.4% and (8.2 +/- 2.0) micro m respectively, which were decreased significantly (P < 0.01) when compared with 2 mmol/L AA treatment group [80.6% and (44.3 +/- 4.0) micro m].
Conclusions:
Acrylamide has significant cytotoxicity and genotoxicity on HaCaT cells. AA-induced DNA damage may be related to the oxidative metabolite(s) of AA through CYP-450.
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