Induction of in situ DNA double-strand breaks and apoptosis by 200 MeV protons and 10 MV X-rays in human tumour cell lines

Int J Radiat Biol. 2011 Jan;87(1):57-70. doi: 10.3109/09553002.2010.518201. Epub 2010 Oct 18.

Abstract

Purpose: To clarify the properties of clinical high-energy protons by comparing with clinical high-energy X-rays.

Materials and methods: Human tumor cell lines, ONS76 and MOLT4, were irradiated with 200 MeV protons or 10 MV X-rays. In situ DNA double-strand breaks (DDSB) induction was evaluated by immunocytochemical staining of phosphorylated histone H2AX (γ-H2AX). Apoptosis was measured by flow-cytometry after staining with Annexin V. The relative biological effectiveness (RBE) was obtained by clonogenic survival assay.

Results: DDSB induction was significantly higher for protons than X-rays with average ratios of 1.28 (ONS76) and 1.59 (MOLT4) at 30 min after irradiation. However the differences became insignificant at 6 h. Also, apoptosis induction in MOLT4 cells was significantly higher for protons than X-rays with an average ratio of 2.13 at 12 h. However, the difference became insignificant at 20 h. RBE values of protons to X-rays at 10% survival were 1.06 ± 0.04 and 1.02 ± 0.15 for ONS76 and MOLT4, respectively.

Conclusions: Cell inactivation may differ according to different timings and/or endpoints. Proton beams demonstrated higher cell inactivation than X-rays in the early phases. These data may facilitate the understanding of the biological properties of clinical proton beams.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / radiation effects*
  • Cell Line, Tumor
  • DNA Breaks, Double-Stranded*
  • Dose-Response Relationship, Radiation
  • Histones / metabolism
  • Humans
  • Medulloblastoma / metabolism
  • Medulloblastoma / pathology
  • Medulloblastoma / radiotherapy
  • Particle Accelerators
  • Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / metabolism
  • Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / pathology
  • Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / radiotherapy
  • Protons*
  • Radiotherapy, High-Energy / instrumentation
  • Relative Biological Effectiveness
  • Synchrotrons
  • Tumor Stem Cell Assay

Substances

  • H2AX protein, human
  • Histones
  • Protons