Ionizing radiation induces long-term senescence in endothelial cells through mitochondrial respiratory complex II dysfunction and superoxide generation

• Published in: Free radical biology & medicine Vol. 108; pp. 750 - 759
• Main Authors: Lafargue, Audrey, Degorre, Charlotte, Corre, Isabelle, Alves-Guerra, Marie-Clotilde, Gaugler, Marie-Hélène, Vallette, François, Pecqueur, Claire, Paris, François
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2017; Elsevier
• Subjects: Apoptosis; Cancer; Cell Line; Cellular Senescence; Electron Transport Complex II - metabolism; Endothelial cell; Endothelial Cells - physiology; Endothelial Cells - radiation effects; Humans; Interleukin-8 - metabolism; Ionizing radiation; Life Sciences; Lysophospholipids - metabolism; Microvessels - pathology; Mitochondria - metabolism; Mitochondrial oxidative stress; Oxidative Stress; Radiation, Ionizing; Senescence; Sphingosine - analogs & derivatives; Sphingosine - metabolism; Superoxide Dismutase - metabolism; Superoxides - metabolism; Tumor Suppressor Protein p53 - metabolism; Mitochondrial oxidative stress; Senescence; Endothelial cell; Ionizing radiation; DDR; ROS; IR; SASP; O/N; mitochondrial oxidative stress; endothelial cell; ionizing radiation
• ISSN: 0891-5849; 1873-4596; 1873-4596
• DOI: 10.1016/j.freeradbiomed.2017.04.019

Ionizing radiation causes oxidative stress, leading to acute and late cellular responses. We previously demonstrated that irradiation of non-proliferating endothelial cells, as observed in normal tissues, induces early apoptosis, which can be inhibited by pretreatment with Sphingosine-1-Phosphate. We now propose to better characterize the long-term radiation response of endothelial cells by studying the molecular pathways associated with senescence and its link with acute apoptosis. First, senescence was validated in irradiated quiescent microvascular HMVEC-L in a dose- and time-dependent manner by SA β-galactosidase staining, p16Ink4a and p21Waf1 expression, pro-inflammatory IL-8 secretion and DNA damage response activation. This premature aging was induced independently of Sphingosine 1-Phosphate treatment, supporting its non-connection with acute IR-induced apoptosis. Then, senescence under these conditions showed persistent activation of p53 pathway and mitochondrial dysfunctions, characterized by O2·- generation, inhibition of respiratory complex II activity and over-expression of SOD2 and GPX1 detoxification enzymes. Senescence was significantly inhibited by treatment with pifithrin–α, a p53 inhibitor, or by MnTBAP, a superoxide dismutase mimetic, validating those molecular actors in IR-induced endothelial cell aging. However, MnTBAP, but not pifithrin–α, was able to limit superoxide generation and to rescue the respiratory complex II activity. Furthermore, MnTBAP was not modulating p53 up-regulation, suggesting that IR-induced senescence in quiescent endothelial cells is provided by at least 2 different pathways dependent of the mitochondrial oxidative stress response and the p53 activation. Further characterization of the actors involved in the respiratory complex II dysfunction will open new pharmacological strategies to modulate late radiation toxicity. [Display omitted] •Ionizing radiation induces long-term senescence in non-proliferating endothelial cells.•Apoptosis inhibition by Sphingosine 1-phosphate does not modulate senescence.•p53 activity, mitochondrial superoxide generation and loss of respiratory complex II activity are observed during senescence.•Superoxide dismutase mimetic or pharmacological inhibition of p53 activation prevents senescence.•SOD mimetic, but not p53 inhibition, blocks superoxide generation and revert respiratory complex II dysfunction.