Single cell time-lapse imaging of focus formation by the DNA damage-response protein 53BP1 after UVC irradiation of human pancreatic cancer cells

Shinji Miwa, Yasunori Tome, Shuya Yano, Yukihiko Hiroshima, Fuminari Uehara, Sumiyuki Mii, Hiroaki Kimura, Katsuhiro Hayashi, Hiroyuki Tsuchiya, Michael Bouvet, Elena V. Efimova, Robert M. Hoffman

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15 Citations (Scopus)

Abstract

We have previously demonstrated that ultraviolet (UV) light treatment is effective against various types of cancer cells expressing fluorescent proteins. In order to further understand the efficacy of UV treatment of cancer cells, we determined the kinetics of focus formation by imaging of a DNA damage-response (DDR) protein after UVC irradiation of human pancreatic cancer cells. A fusion protein consisting of the DDR protein 53BP1 and green fluorescent protein (GFP) (GFP-53BP1) was used as a live-cell imaging marker for cellular response after UVC irradiation. GFP-53BP1 foci were observed after UVC irradiation of MiaPaCa-2 human pancreatic cancer cells. During live-cell imaging, GFP-53BP1 foci were observed in the cells within 15 min after UVC irradiation, and some of the foci remained stable for at least three hours. GFP-53BP1 focus formation was observed in the pancreatic-cancer cells irradiated by 25-200 J/m2 UVC. Our results indicate that an early response to DNA damage caused by UVC irradiation can be visualized by increased GFP-53BP1 focus formation by pancreatic cancer cells.

Original languageEnglish
Pages (from-to)1373-1378
Number of pages6
JournalAnticancer research
Volume33
Issue number4
Publication statusPublished - Apr 2013

Keywords

  • 53BP1
  • DNA damage
  • Focus
  • GFP
  • Pancreatic cancer
  • UVC

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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    Miwa, S., Tome, Y., Yano, S., Hiroshima, Y., Uehara, F., Mii, S., Kimura, H., Hayashi, K., Tsuchiya, H., Bouvet, M., Efimova, E. V., & Hoffman, R. M. (2013). Single cell time-lapse imaging of focus formation by the DNA damage-response protein 53BP1 after UVC irradiation of human pancreatic cancer cells. Anticancer research, 33(4), 1373-1378.