p53和γ-H2AX作为乙醛引起DNA损伤早期生物标记物的实验研究

doi:10.3969/j.issn.1004-616x.2014.02.001

Abstract

Abstract:

OBJECTIVE: The aim of this study was to investigate the changes of biomarkers of DNA damage，total p53，phospho-p53 (p-p53) and phospho-H2AX (p-H2AX or γ-H2AX) following exposure of TK6 cells to acetaldehyde and to explore the roles and the sensitivity of p53，γ-H2AX in DNA damage detection by comparison in the indicators of DNA strand breaks (comet assay). METHODS：TK6 cells were exposed to acetaldehyde with different concentrations of 0.5-20 mmol/L for 20 min，2 and 12 h. Flow cytometry was applied to investigate the changes of relevant biomarkers of DNA damage. Alkaline comet assay was used to detect the indicators of DNA strand breaks (percentage of cells with tails，tail length，percentage of tail DNA，tail moment). RESULTS： After exposure to acetaldehyde for 12 h，a significant increase of γ-H2AX expression was observed at acetaldehyde doses of 15 and 20 mmol/L. A significant dose-dependent increase of p-p53 expression rate was found at acetaldehyde dose between 0.5-7.5 mmol/L. In parallel，a similar trend of expression was observed in total p53，but no significant increase was shown. For the comet assay，significant dose-dependent increases of percentage of cells with tails，tail length，percentage of tail DNA，tail moment were obtained after 12 h exposure acetaldehyde dose between 5.0-12.5 mmol/L. 2 h exposure showed that both total p53 and p-p53 expression rates increased significantly at concentrations of 15 and 20 mmol/L of acetaldehyde，and significant increases of percentage of cells with tails，tail DNA，tail moment accurred at the concentration of 20 mmol/L acetaldehyde，but no response was observed in γ-H2AX. For 20 min exposure，no clear trend for all the three biomarkers and no significant change were shown with the indicators of DNA strand breaks. CONCLUSION： Acetaldehyde could induce increase of p-p53 expression rate，γ-H2AX expression amount，percentage of cells with tails，tail length，percentage of tail DNA，tail moment and total p53 expression rate to a less extent. The biomarker p-p53 may be more sensitive reflecting the DNA damage caused by acetaldehyde than γ-H2AX and indicators of DNA strand breaks.

Key words: acetaldehyde, DNA damage, p53, γ-H2AX, comet assay

ZHOU Xue-xian， Penny Jones， Paul Carmichael， HAO Wei-dong，. Changes of p53 and γ-H2AX of TK6 cells caused by acetaldehyde[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2014.02.001.

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Changes of p53 and γ-H2AX of TK6 cells caused by acetaldehyde

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