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Evaluation of chemical phototoxicity, focusing on phosphorylated histone H2AX.

Ibuki Y, Toyooka T - J. Radiat. Res. (2014)

Bottom Line: Therefore, γ-H2AX is currently attracting attention as a new biomarker for detecting various genotoxic insults.We have determined the toxic impact of various environmental stresses such as chemicals, light and/or their coexposure using γ-H2AX, and found that the γ-H2AX assay exhibited high sensitivity and a low false-positive rate as a detection system of genotoxic potential.In this review, we introduced our recent findings concerning the evaluation of chemical phototoxicity, focusing on γ-H2AX.

View Article: PubMed Central - PubMed

Affiliation: Graduate Division of Nutritional and Environmental Sciences, University of Shizuoka, Shizuoka, Japan ibuki@u-shizuoka-ken.ac.jp.

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Generation of γ-H2AX after treatment with chemicals and UVA. The generation of γ-H2AX 4 h after treatment with chemicals and UVA was analyzed by immunofluorescence staining and western blotting. (A) Images of γ-H2AX foci produced by treatment with 8-MOP and UVA. The graph shows the percentages of γ-H2AX-positive cells (cells having > 10 foci). (B) 8-MOP-dose-dependent (10−9–10−4M) generation of γ-H2AX. Actin was used as a standard for the equal loading of proteins for SDS-PAGE. (C) 6-MC-dose-dependent (10−9–10−4M) generation of γ-H2AX. (D) SDS-dose-dependent (10−5–10−4M) generation of γ-H2AX. The triangles show the limit of the detection. (Edited figure from J Invest Dermatol 2011;131:1313–21 [10].)
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RRU105F5: Generation of γ-H2AX after treatment with chemicals and UVA. The generation of γ-H2AX 4 h after treatment with chemicals and UVA was analyzed by immunofluorescence staining and western blotting. (A) Images of γ-H2AX foci produced by treatment with 8-MOP and UVA. The graph shows the percentages of γ-H2AX-positive cells (cells having > 10 foci). (B) 8-MOP-dose-dependent (10−9–10−4M) generation of γ-H2AX. Actin was used as a standard for the equal loading of proteins for SDS-PAGE. (C) 6-MC-dose-dependent (10−9–10−4M) generation of γ-H2AX. (D) SDS-dose-dependent (10−5–10−4M) generation of γ-H2AX. The triangles show the limit of the detection. (Edited figure from J Invest Dermatol 2011;131:1313–21 [10].)

Mentions: Fig. 5A shows the formation of γ-H2AX 4 h after treatment with 8-MOP under UVA exposure (edited figure from J Invest Dermatol 2011;131:1313–21 [10]). γ-H2AX foci within the nucleus were clearly detected as discrete dots. Cells having >10 foci were significantly increased by the treatment with 8-MOP and UVA, which was 8-MOP-dose-dependent and detectable at 10−9 M. The generation of γ-H2AX was also detected by western blotting from a concentration of 10−9 M, similar to the results of immunofluorescence staining (Fig. 5B). γ-H2AX was also generated after a treatment with the weak phototoxic chemical 6-MC at a concentration of 10−7 M (Fig. 5C). On the other hand, the non-phototoxic compound SDS could not generate γ-H2AX, even at the high doses (Fig. 5D).Fig. 5.


Evaluation of chemical phototoxicity, focusing on phosphorylated histone H2AX.

Ibuki Y, Toyooka T - J. Radiat. Res. (2014)

Generation of γ-H2AX after treatment with chemicals and UVA. The generation of γ-H2AX 4 h after treatment with chemicals and UVA was analyzed by immunofluorescence staining and western blotting. (A) Images of γ-H2AX foci produced by treatment with 8-MOP and UVA. The graph shows the percentages of γ-H2AX-positive cells (cells having > 10 foci). (B) 8-MOP-dose-dependent (10−9–10−4M) generation of γ-H2AX. Actin was used as a standard for the equal loading of proteins for SDS-PAGE. (C) 6-MC-dose-dependent (10−9–10−4M) generation of γ-H2AX. (D) SDS-dose-dependent (10−5–10−4M) generation of γ-H2AX. The triangles show the limit of the detection. (Edited figure from J Invest Dermatol 2011;131:1313–21 [10].)
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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RRU105F5: Generation of γ-H2AX after treatment with chemicals and UVA. The generation of γ-H2AX 4 h after treatment with chemicals and UVA was analyzed by immunofluorescence staining and western blotting. (A) Images of γ-H2AX foci produced by treatment with 8-MOP and UVA. The graph shows the percentages of γ-H2AX-positive cells (cells having > 10 foci). (B) 8-MOP-dose-dependent (10−9–10−4M) generation of γ-H2AX. Actin was used as a standard for the equal loading of proteins for SDS-PAGE. (C) 6-MC-dose-dependent (10−9–10−4M) generation of γ-H2AX. (D) SDS-dose-dependent (10−5–10−4M) generation of γ-H2AX. The triangles show the limit of the detection. (Edited figure from J Invest Dermatol 2011;131:1313–21 [10].)
Mentions: Fig. 5A shows the formation of γ-H2AX 4 h after treatment with 8-MOP under UVA exposure (edited figure from J Invest Dermatol 2011;131:1313–21 [10]). γ-H2AX foci within the nucleus were clearly detected as discrete dots. Cells having >10 foci were significantly increased by the treatment with 8-MOP and UVA, which was 8-MOP-dose-dependent and detectable at 10−9 M. The generation of γ-H2AX was also detected by western blotting from a concentration of 10−9 M, similar to the results of immunofluorescence staining (Fig. 5B). γ-H2AX was also generated after a treatment with the weak phototoxic chemical 6-MC at a concentration of 10−7 M (Fig. 5C). On the other hand, the non-phototoxic compound SDS could not generate γ-H2AX, even at the high doses (Fig. 5D).Fig. 5.

Bottom Line: Therefore, γ-H2AX is currently attracting attention as a new biomarker for detecting various genotoxic insults.We have determined the toxic impact of various environmental stresses such as chemicals, light and/or their coexposure using γ-H2AX, and found that the γ-H2AX assay exhibited high sensitivity and a low false-positive rate as a detection system of genotoxic potential.In this review, we introduced our recent findings concerning the evaluation of chemical phototoxicity, focusing on γ-H2AX.

View Article: PubMed Central - PubMed

Affiliation: Graduate Division of Nutritional and Environmental Sciences, University of Shizuoka, Shizuoka, Japan ibuki@u-shizuoka-ken.ac.jp.

Show MeSH
Related in: MedlinePlus