Expression of Cryptosporidium parvum thioredoxin peroxidase in COS-7 cells confers radioprotection

Exp Parasitol. 2016 Apr:163:8-15. doi: 10.1016/j.exppara.2016.01.012. Epub 2016 Jan 26.

Abstract

Cryptosporidium parvum is one of the most radioresistant organisms identified to date. In a previous study, we found that thioredoxin peroxidase (CpTPx) was significantly upregulated in this species following exposure to high dose (10 kGy) of γ-irradiation. To assess the potential of CpTPx to confer radioprotection in mammalian cells, it was expressed in COS-7 African green monkey kidney cells (CpTPx-COS7). For comparison, the thioredoxin peroxidase of Cryptosporidium muris (CmTPx) was also expressed in these cells (CmTPx-COS7 cells), which has been confirmed to have lesser antioxidant activity than CpTPx in the previous study. Notably, the survival rates of CpTPx-COS7 cells were significantly higher (12-22%) at 72 h after 8 Gy irradiation than CmTPx-COS7 or non-transfected COS-7 (ntCOS-7) counterparts. In addition, CpTPx revealed a 50% of ROS reduction in irradiated CpTPx-COS7 cells, while γ-H2AX DNA damage marker expression was not significantly changed. Furthermore, the amount of apoptosis only increased to about 120% after 2-8 Gy irradiation compared to 200-300% increase observed in ntCOS-7 cells. CmTPx was shown to have antioxidant and DNA damage protection activities; however, these activities were always lower than those of CpTPx. These results suggest that the potent antioxidant and protective activities of CpTPx are well conserved in this cell-based system and that CpTPx contributed to the radioprotection of mammalian cells through its exceptional antioxidant activity.

Keywords: Cryptosporidium parvum; Radioprotection; Thioredoxin peroxidase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antioxidants / metabolism*
  • COS Cells / enzymology*
  • COS Cells / parasitology
  • COS Cells / radiation effects
  • Chlorocebus aethiops
  • Cryptosporidium parvum / enzymology*
  • Cryptosporidium parvum / radiation effects
  • Gamma Rays*
  • Gene Expression Regulation, Enzymologic
  • Microscopy, Confocal
  • Peroxiredoxins / biosynthesis*
  • Peroxiredoxins / genetics
  • Peroxiredoxins / metabolism
  • Reactive Oxygen Species / metabolism
  • Transfection

Substances

  • Antioxidants
  • Reactive Oxygen Species
  • Peroxiredoxins