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Long-wavelength UVA enhances UVB-induced cell death in cultured keratinocytes: DSB formation and suppressed survival pathway

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Abstract

Solar UV radiation consists of both UVA and UVB. The wavelength-specific molecular responses to UV radiation have been studied, but the interaction between UVA and UVB has not been well understood. In this study, we found that long-wavelength UVA, UVA1, augmented UVB-induced cell death, and examined the underlying mechanisms. Human keratinocytes HaCaT were exposed to UVA1, followed by UVB irradiation. Irradiation by UVA1 alone showed no effect on cell survival, whereas the UVA1 pre-irradiation remarkably enhanced UVB-induced cell death. UVA1 delayed the repair of pyrimidine dimers formed by UVB and the accumulation of nucleotide excision repair (NER) proteins to damaged sites. Gap synthesis during NER was also decreased, suggesting that UVA1 delayed NER, and unrepaired pyrimidine dimers and single-strand breaks generated in the process of NER were left behind. Accumulation of this unrepaired DNA damage might have led to the formation of DNA double-strand breaks (DSBs), as was detected using gel electrophoresis analysis and phosphorylated histone H2AX assay. Combined exposure enhanced the ATM–Chk2 signaling pathway, but not the ATR–Chk1 pathway, confirming the enhanced formation of DSBs. Moreover, UVA1 suppressed the UVB-induced phosphorylation of Akt, a survival signal pathway. These results indicated that UVA1 influenced the repair of UVB-induced DNA damage, which resulted in the formation of DSBs and enhanced cell death, suggesting the risk of simultaneous exposure to high doses of UVA1 and UVB.

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Abbreviations

ATM:

Ataxia telangiectasia-mutated

ATR:

ATM and Rad3-related

BER:

Base excision repair

BSA:

Bovine serum albumin

BSFGE:

Biased sinusoidal field gel electrophoresis

CPDs:

Cyclobutane pyrimidine dimers

DSBs:

Double-strand breaks

DMEM:

Dulbecco’s modified Eagle’s medium

EdU:

5-Ethynyl-2′-deoxyuridine

FBS:

Fetal bovine serum

γ-H2AX:

Phosphorylated histone H2AX

NER:

Nucleotide excision repair

8-oxodG:

8-Oxo-7,8-dihydro-2′-deoxyguanosine

PI:

Propidium iodide

PI3K:

Phosphatidylinositol-3 kinase

6-4PPs:

Pyrimidine (6–4) pyrimidone photoproducts

SSBs:

Single strand breaks

ssDNA:

Single-stranded DNA

UV:

Ultraviolet

UVR:

UV radiation

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Acknowledgements

We are grateful to Dr. Vivienne Reeve (University of Sydney) for giving us an excellent suggestion for this research. We also thank Mr. Takuto Okuya for his experimental assistance. This work was supported by Shiseido Female Researcher Science Grant.

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Authors and Affiliations

  1. Graduate Division of Nutritional and Environmental Sciences, University of Shizuoka, 52-1 Yada, Shizuoka, 422-8526, Japan

    Yuko Ibuki, Yukako Komaki & Guang Yang

  2. National Institute of Occupational Safety and Health, Kawasaki, Japan

    Tatsushi Toyooka

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  1. Yuko Ibuki
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  2. Yukako Komaki
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Correspondence to Yuko Ibuki.

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Supplementary Information

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43630_2021_50_MOESM1_ESM.docx

Supplementary file1The wavelength characteristics of UVA1 and UVB radiation, cell survival when irradiation order of UVA1 and UVB is reversed, apoptotic and necrotic cell measurement, the recovery of cell survival by a time interval between UVA1 and UVB exposure, XPA recruitment, quantification of western blotting bands. See https://doi.org/10.1039/x0xx00000x (DOCX 847 kb)

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Ibuki, Y., Komaki, Y., Yang, G. et al. Long-wavelength UVA enhances UVB-induced cell death in cultured keratinocytes: DSB formation and suppressed survival pathway. Photochem Photobiol Sci 20, 639–652 (2021). https://doi.org/10.1007/s43630-021-00050-w

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  • DOI: https://doi.org/10.1007/s43630-021-00050-w

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