Effects of oxygen on the response of mitochondria to X-irradiation and reactive oxygen species-mediated fibroblast activation
Effects of oxygen on the response of mitochondria to X-irradiation and reactive oxygen species-mediated fibroblast activation
复制标题
DOI：
10.1080/09553002.2023.2142980
复制
发表时间：
2022-11
影响因子：
2.6
通讯作者：
T. Shimura;Rina Totani;Hyougo Ogasawara;Keiki Inomata;M. Sasatani;K. Kamiya;A. Ushiyama
中科院分区：
文献类型：
--
作者：
T. Shimura;Rina Totani;Hyougo Ogasawara;Keiki Inomata;M. Sasatani;K. Kamiya;A. Ushiyama
与缺氧或低氧条件下相比，生物体在氧气（O2）存在下对辐射的敏感性增加。在这里，我们研究了O2浓度是否影响线粒体对x射线辐射的反应，这与辐射相关肿瘤中通过成纤维细胞激活形成的肿瘤微环境有关。材料与方法正常人二倍体肺成纤维细胞TIG-3和MRC-5培养过程中，将O2浓度控制在<5%（内环境氧条件）或缺氧水平。使用免疫染色或western blotting评估与线粒体对辐射反应相关的蛋白表达。结果在5%和20% O2条件下，辐射诱导细胞DNA损伤（标记物：γ-H2A组蛋白家族成员X）和线粒体信号（amp活化的蛋白激酶），抑制线粒体代谢活性，产生活性氧。然而，降低O2浓度减轻了辐射对细胞生长、线粒体损伤（parkin）、诱导抗氧化反应（核因子e2相关因子2）和成纤维细胞活化（α-平滑肌肌动蛋白）的影响。在没有氧气的情况下，辐射不会影响本研究中使用的标志物。结论O2浓度影响线粒体对辐射的反应和活性氧介导的成纤维细胞活化。较高的氧浓度增强了辐射对人成纤维细胞线粒体的影响。在体外研究中，由于高氧浓度，可能会高估体内辐射效应。
Abstract Purpose In living organisms, sensitivity to radiation increases in the presence of oxygen (O2) compared with that under anoxic or hypoxic conditions. Here, we investigated whether O2 concentration affected the response of mitochondria to X-rays radiation, which is associated with tumor microenvironment formation via fibroblast activation in radiation-related tumors. Materials and methods O2 concentrations were controlled at <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. Results 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% O2 conditions. However, reducing O2 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 O2. Conclusion O2 concentration affected the response of mitochondria to radiation and reactive oxygen species-mediated fibroblast activation. Higher O2 concentrations enhanced the effects of radiation on mitochondria in human fibroblasts. In vitro studies may overestimate in vivo radiation effects due to high O2 concentrations.