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Real-time observation of irradiated HeLa-cell modified by fluorescent ubiquitination-based cell-cycle indicator using synchrotron X-ray microbeam.

Abstract
Fluorescent ubiquitination-based cell-cycle indicator (FUCCI) human cancer (HeLa) cells (red indicates G1; green, S/G2) were exposed to a synchrotron X-ray microbeam. Cells in either G1 or S/G2 were irradiated selectively according to their colour in the same microscopic field. Time-lapse micrographs of the irradiated cells were acquired for 24 h after irradiation. For fluorescent immunostaining, phosphorylated histone proteins (γ-H2AX) indicated the induction of DNA double-strand breaks. The cell cycle was arrested by irradiation at S/G2. In contrast, cells irradiated at G1 progressed to S/G2. The foci were induced in cells irradiated at both G1 and S/G2, suggesting that the G1-S (or S) checkpoint pathway does not function in HeLa cells due to the fact that the cells are functionally p53 deficient, even though X-ray microbeam irradiation significantly induces double-strand breaks. These results demonstrate that single FUCCI cell exposure and live cell imaging are powerful methods for studying the effects of radiation on the cell cycle.
AuthorsA Narita, K Kaminaga, A Yokoya, M Noguchi, K Kobayashi, N Usami, K Fujii
JournalRadiation protection dosimetry (Radiat Prot Dosimetry) Vol. 166 Issue 1-4 Pg. 192-6 (Sep 2015) ISSN: 1742-3406 [Electronic] England
PMID25870438 (Publication Type: Journal Article)
Copyright© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: [email protected].
Chemical References
  • Fluorescent Dyes
Topics
  • Cell Cycle (radiation effects)
  • DNA Breaks, Double-Stranded (radiation effects)
  • Fluorescent Dyes (analysis)
  • HeLa Cells
  • Humans
  • Image Processing, Computer-Assisted
  • Microscopy, Fluorescence
  • Mitosis (radiation effects)
  • Phosphorylation (radiation effects)
  • Synchrotrons
  • Ubiquitination (radiation effects)
  • X-Rays

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