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Quantification of γH2AX Foci in Response to Ionising Radiation
Published on: April 6, 2010
Phosphorylation of histone H2AX generated by linear alkylbenzene sulfonates and its suppression by UVB exposure
Toru Kubota1, Tatsushi Toyooka, Xiaoxu Zhao
1Institute for Environmental Sciences, University of Shizuoka, Shizuoka, Japan.
Abstract:
We previously demonstrated that the nonionic surfactants, nonylphenol polyethoxylates (NPEOs) induced the phosphorylation of histone H2AX (γ-H2AX), accompanied by DNA double-strand breaks (DSBs), and that exposure to ultraviolet (UV) degraded NPEOs, which sometimes enhanced their DNA-damaging ability. In this study, we showed that linear alkylbenzene sulfonates (LAS), general anion surfactants, also generated DSBs with γ-H2AX, and this ability was attenuated by UVB exposure. In the human breast adenocarcinoma cell line, MCF-7, γ-H2AX was generated in a dose-dependent manner immediately after cells were treated with LAS, and this was attributed to the formation of DSBs and was independent of cell cycle phases. The ability to generate γ-H2AX was markedly reduced in LAS exposed to UVB. HPLC analysis revealed that LAS were a mixture of various alkyl chain lengths, the peaks of which were detected at individual retention times. UVB evenly decreased all peaks of LAS, without migration of peaks to other retention times, which indicated that UVB may degrade the benzene ring of LAS, but did not shorten the alkyl chains. UVB is an important environmental factor in the degradation of LAS exhibiting the ability to induce DSBs, the most serious type of DNA damage.
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