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Imaging DNA Repair After UV Irradiation Damage of Cancer Cells in Gelfoam® Histoculture

  • Shinji Miwa
  • Robert M. Hoffman
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1760)

Abstract

DNA damage repair in response to UVC irradiation was imaged in cancer cells growing in Gelfoam® histoculture. UVC-induced DNA damage repair was imaged with green fluorescent protein (GFP) fused to the DNA damage response (DDR)-related binding protein 53BP1 in MiaPaCa-2 human pancreatic cancer cells. Three-dimensional Gelfoam® histocultures and confocal imaging enabled 53BP1-GFP nuclear foci to be observed within 1 h after UVC irradiation, indicating the onset of DNA damage repair response. Induction of UV-induced 53BP1-GFP focus formation was limited up to a depth of 40 μm in Gelfoam® histoculture of MiaPaCa-2 cells, indicating this was the depth limit of UVC irradiation.

Key words

Cancer cells Gelfoam® Histoculture UV irradiation DNA damage repair 53PB1-GFP Imaging 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Orthopaedic SurgeryKanazawa University Graduate School of MedicineKanazawaJapan
  2. 2.AntiCancer Inc.San DiegoUSA
  3. 3.Department of SurgeryUCSDSan DiegoUSA

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