Garipi 2018 MiPschool Tromso E2
|Phenotyping mitochondrial metabolism in Barrett’s metaplasia-dysplasia-adenocarcinoma sequence: respiratory capacity, extracelular proton flux and ROS production|
Event: MiPschool Tromso-Bergen 2018
With an increasing global incidence of cancer, prevention and therapy is one of the major public health challenges of the 21st century. According to the data of the Global Cancer Observatory, approximately 18.1 million of new cases of cancer are expected in 2018. Esophageal cancer (EC) accounts for 3.2% of the total number of new diagnosed cases. Its severity is reflected in the fact that it is responsible for more than half a million deaths per year and has a 5-year survival rate of only 18% . The main causes for the devastating influence of esophageal cancer are the lack of convenient biological markers and effective treatment options. Metabolism of cancer cells is highly adaptable and expresses great plasticity when it comes to shifting the metabolic pathways with one ultimate goal: to survive and to spread. Numerous advantages are ascribed to such changes, the generation of the energy for life and building blocks for growth being the main one [2,3]. Recently, dysregulated pH emerged as a key player as a survival mechanism in mammalian cancer cells and it is also conferring to them properties of resistance to chemotherapy and invasiveness [4,5]. Whether it is a cause or consequence, this hallmark of most cancer cells contributes to the metabolic shift and to changes in their microenvironment. Also, the alteration in reactive oxygen species (ROS)production has been described as beneficial for cancer cells genesis and survivability. ROS are vital for various cellular processes in both healthy and cancer cells but can also cause oxidative stress in excessive amounts, leading to genetic and functional damage. It is supposed that limiting the Oxidative phosphorylation in cancer cells have protective role in the sense of limiting the amount of ROS generated [3,6]. Our plan is to establish SUIT protocols for functional analysis of respiratory capacities, extracellular proton flux and ROS production in cell lines and human tissue samples of esophageal mucosa and to investigate the influence of different extracellular pH values on the above mentioned parameters. Following human cell lines will be included: radioresistant SCC-25, radiosensitive SCC-090 and HGF cell lines. Experiments are going to be performed using High-Resolution FluoroRespirometry in order to measure mitochondrial respiration, ROS production rate and extracellular proton flux production. Esophageal cell culture models will be used and compared with fresh human esophageal biopsies representing non-cancerous cells/tissue of three different succeeding stages in the Barrett’s metaplasia-dysplasia-adenocarcinoma-sequence. Before starting experiments with experimental cell lines and samples of human tissue, training with HEK239 cells, quality control and proficiency test must be accomplished a proof of competence,skills and research quality. This study is directed towards discovery of new tools for diagnosis and potential targets for a new therapeutic approach in the treatment of esophageal cancer.
Labels: MiParea: Respiration, Patients Pathology: Cancer Stress:Hypoxia Organism: Human Tissue;cell: Endothelial;epithelial;mesothelial cell, Other cell lines Preparation: Permeabilized cells, Permeabilized tissue Enzyme: Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Complex V;ATP synthase, TCA cycle and matrix dehydrogenases, Uncoupling protein
Coupling state: LEAK, ROUTINE, OXPHOS, ET Pathway: CIV, NS, ROX HRR: Oxygraph-2k, O2k-Fluorometer, O2k-Protocol Event: E2
Garipi E(1,2), Iglesias-Gonzalez J(1,2), Gnaiger E(1,2)
- Oroboros Instruments, Innsbruck
- Daniel Swarovski Research Lab, Dept Visceral, Transplant Thoracic Surgery, Medical Univ Innsbruck
Supported by the Marie Skłodowska-Curie PhD Fellowship TRACT.
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