N-Dihydrogalactochitosan Potentiates the Radiosensitivity of Liver Metastatic Tumor Cells Originated from Murine Breast Tumors

Int J Mol Sci. 2019 Nov 8;20(22):5581. doi: 10.3390/ijms20225581.

Abstract

Radiation is a widely used therapeutic method for treating breast cancer. N-dihydrogalactochitosan (GC), a biocompatible immunostimulant, is known to enhance the effects of various treatment modalities in different tumor types. However, whether GC can enhance the radiosensitivity of cancer cells remains to be explored. In this study, triple-negative murine 4T1 breast cancer cells transduced with multi-reporter genes were implanted in immunocompetent Balb/C mice to track, dissect, and identify liver-metastatic 4T1 cells. These cells expressed cancer stem cell (CSC) -related characteristics, including the ability to form spheroids, the expression of the CD44 marker, and the increase of protein stability. We then ex vivo investigated the potential effect of GC on the radiosensitivity of the liver-metastatic 4T1 breast cancer cells and compared the results to those of parental 4T1 cells subjected to the same treatment. The cells were irradiated with increased doses of X-rays with or without GC treatment. Colony formation assays were then performed to determine the survival fractions and radiosensitivity of these cells. We found that GC preferably increased the radiosensitivity of liver-metastatic 4T1 breast cancer cells rather than that of the parental cells. Additionally, the single-cell DNA electrophoresis assay (SCDEA) and γ-H2AX foci assay were performed to assess the level of double-stranded DNA breaks (DSBs). Compared to the parental cells, DNA damage was significantly increased in liver-metastatic 4T1 cells after they were treated with GC plus radiation. Further studies on apoptosis showed that this combination treatment increased the sub-G1 population of cells, but not caspase-3 cleavage, in liver-metastatic breast cancer cells. Taken together, the current data suggest that the synergistic effects of GC and irradiation might be used to enhance the efficacy of radiotherapy in treating metastatic tumors.

Keywords: N-dihydrogalactochitosan (GC), metastatic tumors; cancer stem cells; radiosensitivity; triple-negative breast cancer.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Apoptosis / radiation effects
  • Cell Line, Tumor
  • Cell Separation
  • Cell Survival / drug effects
  • Cell Survival / radiation effects
  • Cell Tracking
  • Chitosan / pharmacology*
  • DNA Damage
  • Female
  • Histones / metabolism
  • Liver Neoplasms / pathology
  • Liver Neoplasms / radiotherapy*
  • Liver Neoplasms / secondary*
  • Mammary Neoplasms, Animal / pathology*
  • Mice, Inbred BALB C
  • Molecular Imaging
  • Neoplastic Stem Cells / drug effects
  • Neoplastic Stem Cells / pathology
  • Neoplastic Stem Cells / radiation effects
  • Radiation Tolerance / drug effects*
  • Radiation Tolerance / radiation effects
  • Radiation, Ionizing
  • X-Rays

Substances

  • Histones
  • gamma-H2AX protein, mouse
  • Chitosan