γH2AX, an Accurate Marker That Analyzes UV Genotoxic Effects on Human Keratinocytes and on Human Skin

Barnes, Larry D.; Dumas, Marc; Juan, Mylèbne; Noblesse, Emmanuelle; Tesniere, Anne; Schnebert, Sylvianne; Guillot, B.; Molès, Jean‐Pierre

doi:10.1111/j.1751-1097.2010.00744.x

Cited by 31 publications

(32 citation statements)

References 33 publications

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“…γ-H2AX, which has been identified as an early event after DSBs formation, is considered the most sensitive assay for DSBs detection [29]. In our study, we found that irradiation of UVB resulted in an increased expression ofγH2AX.…”

Section: Discussionsupporting

confidence: 57%

The Protective Effect of Baicalin against UVB Irradiation Induced Photoaging: An In Vitro and In Vivo Study

Zhang

Yin

et al. 2014

PLoS ONE

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ObjectiveThis study was aimed to evaluate the anti-photoaging effects of baicalin on Ultraviolet B (UVB)-induced photoaging in the dorsal skin of hairless mice and premature senescence in human dermal fibroblasts.MethodsWe established in vivo and in vitro photoaging models by repeated exposures to UVB irradiation. By HE staining, masson staining, immunohistostaing and real-time RT-PCR, we analyzed epidermal thickness, collagen expression and the mRNA and protein levels of type I collagen, type III collagen, interstitial collagenase (MMP-1 and MMP-3) in UVB exposed dorsal mice skin. The aging condition in human dermal fibroblasts was determined by senescence-associated β-galactosidase (SA-β-gal) staining. Cell viability was determined using the Cell Counting Kit-8 (CCK-8). The G1 phase cell growth arrest was analyzed by flow cytometry. The senescence-related protein levels of p16INK-4a, p21WAF-1, and p53 and protein levels of phosphorylated histone H2AX were estimated by Western blotting.ResultsTopically application of baicalin treatment reduced UVB-induced epidermal thickening of mouse skin and also result in an increase in the production of collagen I and III, and a decrease in the expression of MMP-1 and MMP-3. Compared with the UVB-irradiated group, we found that the irradiated fibroblasts additionally treated with baicalin demonstrated a decrease in the expression of SA-β-gal, a increase in the cell viability, a decrease in the G1 phase cell proportion, a downregulation in the level of senescence-associated and γ-H2AX proteins. However, Baicalin had no difference in the normal fibroblasts without UVB irradiation and long-term Baicalin incubation of UVB-SIPS fibroblasts gave no effects on the cell proliferation.ConclusionsTaken together, these results suggest that baicalin significantly antagonizes photoaging induced by UVB in vivo and in vitro, indicating the potential of baicalin application for anti-photoaging treatment.

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Section: Discussionsupporting

confidence: 57%

The Protective Effect of Baicalin against UVB Irradiation Induced Photoaging: An In Vitro and In Vivo Study

Zhang

Yin

et al. 2014

PLoS ONE

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“…3A, P = 0.0001). Phosphohistone H2A.X has been shown to be a surrogate marker for dsDNA breaks (DSB) induced by UVB (26). Following UVB exposure, the levels of phosphohistone H2A.X peaked at 6 hours in both vehicle-and CPSI-1306–treated epidermis, following which the levels returned to nearly baseline levels.…”

Section: Resultsmentioning

confidence: 99%

MIF Antagonist (CPSI-1306) Protects against UVB-Induced Squamous Cell Carcinoma

Nagarajan

Tober

Riggenbach

et al. 2014

Molecular Cancer Research

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Macrophage migration inhibitory factor (MIF) is a homotrimeric proinflammatory cytokine implicated in chronic inflammatory diseases and malignancies, including cutaneous squamous cell carcinomas (SCC). To determine whether MIF inhibition could reduce UVB light–induced inflammation and squamous carcinogenesis, a small-molecule MIF inhibitor (CPSI-1306) was utilized that disrupts homotrimerization. To examine the effect of CPSI-1306 on acute UVB-induced skin changes, Skh-1 hairless mice were systemically treated with CPSI-1306 for 5 days before UVB exposure. In addition to decreasing skin thickness and myeloperoxidase (MPO) activity, CPSI-1306 pretreatment increased keratinocyte apoptosis and p53 expression, decreased proliferation and phosphohistone variant H2AX (γ-H2AX), and enhanced repair of cyclobutane pyrimidine dimers. To examine the effect of CPSI-1306 on squamous carcinogenesis, mice were exposed to UVB for 10 weeks, followed by CPSI-1306 treatment for 8 weeks. CPSI-1306 dramatically decreased the density of UVB-associated p53 foci in non–tumor-bearing skin while simultaneously decreasing the epidermal Ki67 proliferation index. In addition to slowing the rate of tumor development, CPSI-1306 decreased the average tumor burden per mouse. Although CPSI-1306–treated mice developed only papillomas, nearly a third of papillomas in vehicle-treated mice progressed to microinvasive SCC. Thus, MIF inhibition is a promising strategy for prevention of the deleterious cutaneous effects of acute and chronic UVB exposure.

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“…Less clear is the functional role of γH2Ax during nucleotide excision repair (NER) and arrest of DNA replication following large adduct damage, such as UV light-induced photoproducts and DNA crosslinks where DSBs are presumably not initially formed (6)(7)(8)(9)(10)(11). Several distinct distributions of γH2Ax occur in UV damaged cells (12)(13)(14)(15)(16)(17).…”

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confidence: 99%

Functional relevance of the histone γH2Ax in the response to DNA damaging agents

Revet

Feeney²,

Bruguera³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

116

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The phosphorylation of H2Ax on its S139 site, γH2Ax, is important during DNA double-strand repair and is considered necessary for assembly of repair complexes, but its functional role after other kinds of DNA damage is less clear. We have measured the survival of isogenic mouse cell lines with the H2Ax gene knocked out, and replaced with wild-type or mutant (S139A) H2Ax genes, exposed to a range of agents with varied mechanisms of DNA damage. Knockout and mutant cells were sensitive to γ-rays, etoposide, temozolamide, and endogenously generated reactive oxygen species, each of which can include double-strand breaks among their spectra of DNA lesions. The absence or mutation of H2Ax had no influence on sensitivity to cisplatin or mitomycin C. Although UV light induced the highest levels of γH2Ax, mutation of S139 had no influence on UV sensitivity or the UV DNA damage response. Complete loss of H2Ax reduced the survival of cells exposed to UV light and reduced pChk1 induction, suggesting that sites other than S139 may impact the ATR-pChk1 pathway. The relative intensity of γH2Ax measured in Western blots in wild-type cells did not correlate with the functional importance of γH2Ax. The use of γH2Ax as a general biomarker of DNA damage is therefore potentially misleading because it is not an unambiguous indicator of double-strand breaks, and a significant fraction of DNA repair, especially involving nucleotide excision or crosslink repair, can occur without functional involvement of γH2Ax.alkylation | rotenone | caffeine | poly(ADP-ribose) polymerase | veliparib

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γH2AX, an Accurate Marker That Analyzes UV Genotoxic Effects on Human Keratinocytes and on Human Skin

Cited by 31 publications

References 33 publications

The Protective Effect of Baicalin against UVB Irradiation Induced Photoaging: An In Vitro and In Vivo Study

The Protective Effect of Baicalin against UVB Irradiation Induced Photoaging: An In Vitro and In Vivo Study

MIF Antagonist (CPSI-1306) Protects against UVB-Induced Squamous Cell Carcinoma

Functional relevance of the histone γH2Ax in the response to DNA damaging agents

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