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

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


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 


  1. 1.
    Miwa S, Yano S, Hiroshima Y, Tome Y, Uehara F, Mii S, Efimove EV, Kimura H, Hayashi K, Tsuchiya H, Hoffman RM (2013) Imaging UVC-induced DNA damage response in models of minimal cancer. J Cell Biochem 114:2493–2499CrossRefPubMedGoogle Scholar
  2. 2.
    Efimova EV, Mauceri HJ, Golden DW, Labay E, Bindokas VP, Darga TE, Chakraborty C, BarretoAndrade JC, Crawley C, Sutton HG, Kron SJ, Weichselbaum RR (2010) Poly (ADP-ribose) polymerase inhibitor induces accelerated senescence in irradiated breast cancer cells and tumors. Cancer Res 70:6277–6282CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Leighton JA (1951) Sponge matrix method for tissue culture; formation of organized aggregates of cells in vitro. J Natl Cancer Inst 12:545–561PubMedGoogle Scholar
  4. 4.
    Freeman AE, Hoffman RM (1986) In vivo-like growth of human tumors in vitro. Proc Natl Acad Sci U S A 83:2694–2698CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Vescio RA, Redfern CH, Nelson TJ, Ugoretz S, Stern PH, Hoffman RM (1987) In vivo-like drug responses of human tumors growing in three-dimensional gel-supported, primary culture. Proc Natl Acad Sci U S A 84:5029–5033CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Hoffman RM (2010. Published Online) Histocultures and their use. In: Encyclopedia of life sciences. Wiley, Chichester. Google Scholar

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

Personalised recommendations