- Email: [email protected]
Glutathione and the switch of aerobic metabolism collaborate for multi-drug resistance of neuroblastoma
D. Rossin et al. / Free Radical Biology and Medicine 108 (2017) S18–S107
(BCKDH).Blockage in BCKDH activity leads to classic Maple Syrup Urine Disease (MSUD).To study the roles of BCAAs, we used cells with a single gene defect in BCKDH as a cellular model.We studied fibroblasts from four unrelated patients with null mutations in BCKDH and from controls.Fibroblasts from patients showed 2-fold increase in mitochondrial superoxide levels and 1.5-fold increase in protein carbonylation levels respect to controls.No changes in SOD2 protein levels were detected, indicating an increase production of mitochondrial superoxide and not an increase detoxification.Eleven proteins related to oxidative stress were differentially regulated in MSUD (p-value 0.05).Including up-regulated peroxiredoxin-4(PRDX4) and protein disulphide-isomerase(P4HB), as well as down-regulated prostaglandin G/H synthase 1 (PTGS1), also known as cyclooxygenase-1.These results correlate specific proteins to known effects of BCAAs in oxidative stress and shed light in the pharmacological mechanisms of BCAAs supplementation. E-mail address: [email protected] (P. Fernandez Guerra) Acknowledgements
Supported by Department of Clinical Medicine and Faculty of Health, Aarhus University, and Aarhus University Research Foundation (AUFF) Aarhus, Denmark. The John and Birthe Meyer Foundation. Fundacion Ramon Areces, grant number: CIVP16A1853, Spain. http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.234
P-150
Chronic UV irradiation induced oxidative stress in the skin of diabetic hairless mice Maria Giakoumaki, Maria Kyriazi, Aggeliki Dimakopoulou, Vasiliki Anagnostou, Marina Karasmani, Andreas Grigoropoulos, Eleni Deli, Nick Andreou, Michail Rallis National University of Athens, Greece Keywords: Skin; inflammation; diabetes; antioxidants; oxidative stress; ultraviolet radiation
Diabetic mellitus induces many pathophysiological changes in skin. The effect of UV light on chronic exposure in diabetic and normal skin was investigated. Hairless mice skin, diabetic (D) and non-diabetic (ND), were exposed to UVA and UVB radiation 3 times per week for 18 weeks. The irradiation dose was equal to 0.75 M.E.D during the first week and increased by 25% each week until the maximal dose was 3.5 M.E.D. Diabetes was induced by streptozotocin injection. Stratum corneum hydration of D mice was significantly lower than ND (p o0.01). The transepidermal water loss of D mice was less than ND mice due to the dryness of the skin of D mice. The elasticity of ND mice's skin was significantly higher (p o0.05). Skin sebum in the D mice was much lower in relation to ND (p o0.05). Skin of D mice is more pigmented and thinner than this of ND. Hydrophilic antioxidants and oxidative stress in both SC and total skin of D and ND mice is in process of evaluation. This results suggest that D mice presented more severe inflammation, photoageing and hyperpigmentation after chronic UV irradiation.
S69
http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.235
P-151
Glutathione and the switch of aerobic metabolism collaborate for multi-drug resistance of neuroblastoma Cinzia Domenicotti 1, Andrea Speciale 1, Ombretta Garbarino 1, Alberto Izzotti 2,3, Daniela Fenoglio 4, Silvia Ravera 5, Alessandra Pulliero 2, Mario Passalacqua 1, Nicola Traverso 1, Maria Adelaide Pronzato 1, Barbara Marengo 1 1
Department of Experimental Medicine, University of Genova, Italy Department of Health Sciences, University of Genova, Italy 3 2IRCCS AOU San Martino IST Genova, Italy 4 Center of Excellence for Biomedical Research, Department of Internal Medicine, University of Genova, Italy 5 Department of Pharmacy, University of Genova, Italy 2
Keywords: GSH; metabolism
neuroblastoma;
BSO;
chemoresistance;
The availability of antioxidants is recognized as one of the critical factors able to make cancer cells resistant to chemotherapy. In this context, it has been demonstrated that many chemoresistant cancers display high levels of glutathione (GSH) and consequently, its depletion by L-buthionine sulfoximine (BSO), has been proposed as a chemosensitizing therapy. To investigate the role of GSH and of tumor metabolism in multi-drug resistance (MDR), HTLA-230 neuroblastoma cells were chronically treated with etoposide at a concentration that in vitro mimics the clinically-used dose. The selected cells (HTLA-Chr) were highly tumorigenic and acquired MDR, becoming less sensitive to etoposide or doxorubicin compared to parental cells. Moreover, HTLA-Chr cells, while having an efficient aerobic metabolism, owing to a favourable P/O ratio and a decreased formation of lactic acid, were also characterized by an up-regulation of catalase and higher levels of GSH. BSO treatment of HTLA-Chr cells markedly reduced their tumorigenicity that was, instead, enhanced by N-Acetylcysteine, able to promote GSH synthesis. Collectively, our results show that GSH and the switch of aerobic metabolism collaborate for the acquisition of MDR, providing pre-clinical evidence that may drive future therapeutic approaches for sensitizing neuroblastoma to conventional therapies. E-mail address: [email protected] (C. Domenicotti) Acknowledgements
Supported by Genoa University http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.236
P-152
Skin Inflammation and Oxidative Stress Michail Rallis, Maria Kyriazi, Georgios Th. Papaioannou, Andreas Vitsos, Sotirios Liakos, Styliani Daskalaki, Angeliki Dimakopoulou, Maria Giakoumaki, Nikolaos Panayiotis Andreou, Heleni Deli, Vasiliki Anagnostou, Marianna Papageorgiadi, Marina Karasmani National and Kapodistrian University of Athens, Greece
E-mail address: [email protected] (M. Giakoumaki)
aerobic
(BCKDH).Blockage in BCKDH activity leads to classic Maple Syrup Urine Disease (MSUD).To study the roles of BCAAs, we used cells with a single gene defect in BCKDH as a cellular model.We studied fibroblasts from four unrelated patients with null mutations in BCKDH and from controls.Fibroblasts from patients showed 2-fold increase in mitochondrial superoxide levels and 1.5-fold increase in protein carbonylation levels respect to controls.No changes in SOD2 protein levels were detected, indicating an increase production of mitochondrial superoxide and not an increase detoxification.Eleven proteins related to oxidative stress were differentially regulated in MSUD (p-value 0.05).Including up-regulated peroxiredoxin-4(PRDX4) and protein disulphide-isomerase(P4HB), as well as down-regulated prostaglandin G/H synthase 1 (PTGS1), also known as cyclooxygenase-1.These results correlate specific proteins to known effects of BCAAs in oxidative stress and shed light in the pharmacological mechanisms of BCAAs supplementation. E-mail address: [email protected] (P. Fernandez Guerra) Acknowledgements
Supported by Department of Clinical Medicine and Faculty of Health, Aarhus University, and Aarhus University Research Foundation (AUFF) Aarhus, Denmark. The John and Birthe Meyer Foundation. Fundacion Ramon Areces, grant number: CIVP16A1853, Spain. http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.234
P-150
Chronic UV irradiation induced oxidative stress in the skin of diabetic hairless mice Maria Giakoumaki, Maria Kyriazi, Aggeliki Dimakopoulou, Vasiliki Anagnostou, Marina Karasmani, Andreas Grigoropoulos, Eleni Deli, Nick Andreou, Michail Rallis National University of Athens, Greece Keywords: Skin; inflammation; diabetes; antioxidants; oxidative stress; ultraviolet radiation
Diabetic mellitus induces many pathophysiological changes in skin. The effect of UV light on chronic exposure in diabetic and normal skin was investigated. Hairless mice skin, diabetic (D) and non-diabetic (ND), were exposed to UVA and UVB radiation 3 times per week for 18 weeks. The irradiation dose was equal to 0.75 M.E.D during the first week and increased by 25% each week until the maximal dose was 3.5 M.E.D. Diabetes was induced by streptozotocin injection. Stratum corneum hydration of D mice was significantly lower than ND (p o0.01). The transepidermal water loss of D mice was less than ND mice due to the dryness of the skin of D mice. The elasticity of ND mice's skin was significantly higher (p o0.05). Skin sebum in the D mice was much lower in relation to ND (p o0.05). Skin of D mice is more pigmented and thinner than this of ND. Hydrophilic antioxidants and oxidative stress in both SC and total skin of D and ND mice is in process of evaluation. This results suggest that D mice presented more severe inflammation, photoageing and hyperpigmentation after chronic UV irradiation.
S69
http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.235
P-151
Glutathione and the switch of aerobic metabolism collaborate for multi-drug resistance of neuroblastoma Cinzia Domenicotti 1, Andrea Speciale 1, Ombretta Garbarino 1, Alberto Izzotti 2,3, Daniela Fenoglio 4, Silvia Ravera 5, Alessandra Pulliero 2, Mario Passalacqua 1, Nicola Traverso 1, Maria Adelaide Pronzato 1, Barbara Marengo 1 1
Department of Experimental Medicine, University of Genova, Italy Department of Health Sciences, University of Genova, Italy 3 2IRCCS AOU San Martino IST Genova, Italy 4 Center of Excellence for Biomedical Research, Department of Internal Medicine, University of Genova, Italy 5 Department of Pharmacy, University of Genova, Italy 2
Keywords: GSH; metabolism
neuroblastoma;
BSO;
chemoresistance;
The availability of antioxidants is recognized as one of the critical factors able to make cancer cells resistant to chemotherapy. In this context, it has been demonstrated that many chemoresistant cancers display high levels of glutathione (GSH) and consequently, its depletion by L-buthionine sulfoximine (BSO), has been proposed as a chemosensitizing therapy. To investigate the role of GSH and of tumor metabolism in multi-drug resistance (MDR), HTLA-230 neuroblastoma cells were chronically treated with etoposide at a concentration that in vitro mimics the clinically-used dose. The selected cells (HTLA-Chr) were highly tumorigenic and acquired MDR, becoming less sensitive to etoposide or doxorubicin compared to parental cells. Moreover, HTLA-Chr cells, while having an efficient aerobic metabolism, owing to a favourable P/O ratio and a decreased formation of lactic acid, were also characterized by an up-regulation of catalase and higher levels of GSH. BSO treatment of HTLA-Chr cells markedly reduced their tumorigenicity that was, instead, enhanced by N-Acetylcysteine, able to promote GSH synthesis. Collectively, our results show that GSH and the switch of aerobic metabolism collaborate for the acquisition of MDR, providing pre-clinical evidence that may drive future therapeutic approaches for sensitizing neuroblastoma to conventional therapies. E-mail address: [email protected] (C. Domenicotti) Acknowledgements
Supported by Genoa University http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.236
P-152
Skin Inflammation and Oxidative Stress Michail Rallis, Maria Kyriazi, Georgios Th. Papaioannou, Andreas Vitsos, Sotirios Liakos, Styliani Daskalaki, Angeliki Dimakopoulou, Maria Giakoumaki, Nikolaos Panayiotis Andreou, Heleni Deli, Vasiliki Anagnostou, Marianna Papageorgiadi, Marina Karasmani National and Kapodistrian University of Athens, Greece
E-mail address: [email protected] (M. Giakoumaki)
aerobic