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In living organisms, sensitivity to radiation increases in the presence of oxygen (O 2 ) compared with that under anoxic or hypoxic conditions. Here, we investigated whether O 2  concentration affected the response of mitochondria to X-rays radiation, which is associated with tumor microenvironment formation via fibroblast activation in radiation-related tumors. O 2  concentrations were controlled at &lt;5% (internal environmental oxygen condition) or anoxic levels during culture of normal human diploid lung fibroblasts TIG-3 and MRC-5. Protein expression associated with the response of mitochondria to radiation was assessed using immunostaining or western blotting. Induction of DNA damage (marker: γ-H2A histone family member X) and mitochondrial signaling (AMP-activated protein kinase), suppression of mitochondrial metabolic activity, and generation of reactive oxygen species occurred with radiation in cells cultured under 5% and 20% O 2  conditions. However, reducing O 2  concentration mitigated the effects of radiation on cell growth, mitochondrial damage (parkin), induction of antioxidant responses (nuclear factor E2-related factor 2), and fibroblast activation ( α -smooth muscle actin). Radiation did not affect the markers used in this study in the absence of O 2 . O 2  concentration affected the response of mitochondria to radiation and reactive oxygen species-mediated fibroblast activation. Higher O 2  concentrations enhanced the effects of radiation on mitochondria in human fibroblasts. In vitro studies may overestimate in vivo radiation effects due to high O 2  concentrations.

Effects of oxygen on the response of mitochondria to X-irradiation and reactive oxygen species-mediated fibroblast activation