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Difference between revisions of "Chang 2018 Life Sciences Meeting 2018 Innsbruck AT"

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== Affiliations and support ==
== Affiliations ==
Shao Chiang Chang(1), Timea Komlodi(1), Jozsef Dudas(3), Erich Gnaiger(1,2)
Shao Chiang Chang(1), Timea Komlodi(1), Jozsef Dudas(3), Erich Gnaiger(1,2)



Revision as of 16:21, 22 March 2018

pH dependence of mitochondrial respiration and H2O2 production in oral cancer cells – a pilot study

Link: Life Sciences Meeting 2018 Innsbruck AT

Chang SC, Komlodi T, Dudas J, Gnaiger E (2018)

Event: Life Sciences Meeting 2018 Innsbruck AT

Introduction: Cancers are characterized by a high metabolic plasticity resulting from mutations and the selection of metabolic phenotypes. Metabolic transformations involve mitochondrial adaptation or mitochondrial dysfunction. Metabolism and mitochondria have become focal targets for anticancer therapy. In the progression of tumor, cancer cells upregulate glucose uptake and glycolysis. The different environmental and intracellular pH due to the metabolic reprogramming can change the behavior of the cells. Therefore, we compared the effects of pH on mitochondrial respiration and H2O2 production in different cell lines.

Methods: We measured respiration and H2O2 production in permeabilized human embryonic kidney cells (HEK239T), human gingival fibroblasts (HGF) and human oral squamous carcinoma cancer cells (SCC25) in the pH range from 6.6 – 7.5 using High-Resolution FluoRespirometry including the Amplex UltraRed assay. We used substrate-uncoupler-inhibitor titration protocols to access mitochondrial respiration in different coupling and pathway control states, activating the succinate (S)-, NADH- (N) and fatty acid (F)- pathways separately or in combination.

Results: With increasing pH, H2O2 flux decreased in HEK239T cells in the S- and N-pathways, while O2 flux was significantly increased. A similar result was observed in HGF. Conversely, the SCC25 cells showed an opposite trend. In these cancer cells, electron transfer pathways and Complex IV activity were depressed at high pH. HGF and SCC25 showed a higher sensitivity to intracellular pH compared to HEK293T cells.

Conclusion: SCC25 cancer cells showed a response of respiration to changes in pH that was opposite compared to normal HGF and HEK293T cells. The sensitivity of mitochondrial respiration to modulations of intracellular pH may have implications on chemotherapy sensitivity. Therefore, the experimental design evaluated in the present study will be applied in radioresistant SCC25 cells and radiochemotherapy sensitive SCC-090 cells.


β€’ Bioblast editor: Chang SC β€’ O2k-Network Lab: AT Innsbruck Oroboros


Labels: MiParea: Respiration  Pathology: Cancer 

Organism: Human  Tissue;cell: HEK  Preparation: Permeabilized cells 


Coupling state: LEAK, ROUTINE, OXPHOS, ET  Pathway: F, N, S, Gp, CIV, NS, Other combinations, ROX  HRR: Oxygraph-2k, O2k-FluoRespirometer"O2k-FluoRespirometer" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property. 


Affiliations

Shao Chiang Chang(1), Timea Komlodi(1), Jozsef Dudas(3), Erich Gnaiger(1,2)

  1. Oroboros Instruments, Innsbruck, Austria
  2. Department Visceral, Thoracic Surgery, Medical University Innsbruck, Innsbruck, Austria
  3. Molecularbiological Oncological Research Group, Department Otorhinolaryngology and Head & Neck Surgery, Medical University Innsbruck, Innsbruck, Austria