title: Oxidative phosphorylation and mitochondrial function differ between human prostate tissue and cultured cells.
abstract: Altered mitochondrial metabolism plays a pivotal role in the development and progression of various diseases, including cancer. Cell lines are frequently used as models to study mitochondrial (dys)function, but little is known about their mitochondrial respiration and metabolic properties in comparison to the primary tissue of origin. We have developed a method for assessment of oxidative phosphorylation in prostate tissue samples of only 2 mg wet weight using high-resolution respirometry. Reliable protocols were established to investigate the respiratory activity of different segments of the mitochondrial electron transfer system (ETS) in mechanically permeabilized tissue biopsies. Additionally, the widely used immortalized prostate epithelial and fibroblast cell lines, RWPE1 and NAF, representing the major cell types in prostate tissue, were analyzed and compared to the tissue of origin. Our results show that mechanical treatment without chemical permeabilization agents or sample processing constitutes a reliable preparation method for OXPHOS analysis in small amounts of prostatic tissue typically obtained by prostate biopsy. The cell lines represented the bioenergetic properties of fresh tissue to a limited extent only. Particularly, tissue showed a higher oxidative capacity with succinate and glutamate, whereas pyruvate was a substrate supporting significantly higher respiratory activities in cell lines. Several fold higher zinc levels measured in tissue compared to cells confirmed the role of aconitase for prostate-specific metabolism in agreement with observed respiratory properties. In conclusion, combining the flexibility of cell culture models and tissue samples for respirometric analysis are powerful tools for investigation of mitochondrial function and tissue-specific metabolism.
authors.affiliation: Division of Genetic Epidemiology, Department of Medical Genetics, Molecular and Clinical Pharmacology, Medical University of Innsbruck, Austria.
authors.forename: Bernd
authors.initials: B
authors.lastname: Schöpf
authors.affiliation: Division of Experimental Urology, Department of Urology, Medical University of Innsbruck, Austria.|Department of Pathology, Medical University of Innsbruck, Austria.
authors.forename: Georg
authors.initials: G
authors.lastname: Schäfer
authors.affiliation: Division of Experimental Urology, Department of Urology, Medical University of Innsbruck, Austria.
authors.forename: Anja
authors.initials: A
authors.lastname: Weber
authors.affiliation: Biocenter, Section for Clinical Biochemistry, Medical University of Innsbruck, Austria.
authors.forename: Heribert
authors.initials: H
authors.lastname: Talasz
authors.affiliation: Division of Experimental Urology, Department of Urology, Medical University of Innsbruck, Austria.
authors.forename: Iris E
authors.initials: IE
authors.lastname: Eder
authors.affiliation: Division of Experimental Urology, Department of Urology, Medical University of Innsbruck, Austria.
authors.forename: Helmut
authors.initials: H
authors.lastname: Klocker
authors.affiliation: Department of General and Transplant Surgery, D. Swarovski Research Laboratory, Medical University of Innsbruck, Austria.
authors.forename: Erich
authors.initials: E
authors.lastname: Gnaiger
mesh_terms: D002460:Cell Line; D019069:Cell Respiration / Q000235:genetics; D002478:Cells, Cultured / Q000378:metabolism; D004579:Electron Transport; D004734:Energy Metabolism; D005347:Fibroblasts / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D008297:Male; D008931:Mitochondria, Muscle / Q000378:metabolism* / Q000473:pathology; D010085:Oxidative Phosphorylation; D010101:Oxygen Consumption / Q000235:genetics; D011467:Prostate / Q000378:metabolism* / Q000473:pathology; D019289:Pyruvic Acid / Q000378:metabolism; D019802:Succinic Acid / Q000378:metabolism
chemical_list: D018698:Glutamic Acid; D019289:Pyruvic Acid; D019802:Succinic Acid
keywords: electron transfer system; high-resolution respirometry; mitochondria; oxidative phosphorylation; prostate metabolism