Metabolic remodeling precedes mitochondrial outer membrane permeabilization in human glioma xenograft cells.

Glioma cancer cells adapt to changing microenvironment and shift from mitochondrial oxidative phosphorylation to aerobic glycolysis for their metabolic needs irrespective of oxygen availability. In the present study, we show that silencing MMP-9 in combination with uPAR/cathepsin B switch the glycolytic metabolism of glioma cells to oxidative phosphorylation (OXPHOS) and generate reactive oxygen species (ROS) to predispose glioma cells to mitochondrial outer membrane permeabilization. shRNA for MMP-9 and uPAR (pMU) as well as shRNA for MMP-9 and cathepsin B (pMC) activated complexes of mitochondria involved in OXPHOS and inhibited glycolytic hexokinase expression. The decreased interaction of hexokinase 2 with mitochondria in the treated cells indicated the inhibition of glycolysis activation. Overexpression of Akt reversed the pMU- and pMC-mediated OXPHOS to glycolysis switch. The OXPHOS un-coupler oligomycin A altered the expression levels of the Bcl-2 family of proteins; treatment with pMU or pMC reversed this effect and induced mitochondrial outer membrane permeabilization. In addition, our results show changes in mitochondrial pore transition to release cytochrome c due to changes in the VDAC-Bcl-XL and BAX-BAK interaction with pMU and pMC treatments. Taken together, our results suggest that pMU and pMC treatments switch glioma cells from the glycolytic to the OXPHOS pathway through an inhibitory effect on Akt, ROS induction and an increase of cytosolic cytochrome c accumulation. These results demonstrate the potential of pMU and pMC as therapeutic candidates for the treatment of glioma.

[1]  A. Guha,et al.  Developmental profile and regulation of the glycolytic enzyme hexokinase 2 in normal brain and glioblastoma multiforme , 2011, Neurobiology of Disease.

[2]  A. Guha,et al.  Targeting Metabolic Remodeling in Glioblastoma Multiforme , 2010, Oncotarget.

[3]  R. Hamanaka,et al.  Mitochondrial reactive oxygen species regulate cellular signaling and dictate biological outcomes. , 2010, Trends in biochemical sciences.

[4]  C. Gondi,et al.  Urokinase Plasminogen Activator Receptor and/or Matrix Metalloproteinase-9 Inhibition Induces Apoptosis Signaling through Lipid Rafts in Glioblastoma Xenograft Cells , 2010, Molecular Cancer Therapeutics.

[5]  C. Gondi,et al.  MMP-9, uPAR and Cathepsin B Silencing Downregulate Integrins in Human Glioma Xenograft Cells In Vitro and In Vivo in Nude Mice , 2010, PloS one.

[6]  Yongqiang Chen,et al.  Regulation of autophagy by reactive oxygen species (ROS): implications for cancer progression and treatment. , 2009, Antioxidants & redox signaling.

[7]  N. Denko,et al.  Hypoxia, HIF1 and glucose metabolism in the solid tumour , 2008, Nature Reviews Cancer.

[8]  A. Bergenheim,et al.  Glucose metabolites, glutamate and glycerol in malignant glioma tumours during radiotherapy , 2008, Journal of Neuro-Oncology.

[9]  R. Meacham,et al.  Oxidative stress is inherent in prostate cancer cells and is required for aggressive phenotype. , 2008, Cancer research.

[10]  N. Huseby,et al.  The Role of Reactive Oxygen Species in Integrin and Matrix Metalloproteinase Expression and Function , 2008, Connective tissue research.

[11]  Lewis C. Cantley,et al.  AKT/PKB Signaling: Navigating Downstream , 2007, Cell.

[12]  M. Los,et al.  Selected technologies to control genes and their products for experimental and clinical purposes , 2007, Archivum Immunologiae et Therapiae Experimentalis.

[13]  G. Semenza,et al.  HIF-1 inhibits mitochondrial biogenesis and cellular respiration in VHL-deficient renal cell carcinoma by repression of C-MYC activity. , 2007, Cancer cell.

[14]  Richard D. Vaughan-Jones,et al.  Regulation of tumor pH and the role of carbonic anhydrase 9 , 2007, Cancer and Metastasis Reviews.

[15]  S. Cory,et al.  The Bcl-2 apoptotic switch in cancer development and therapy , 2007, Oncogene.

[16]  E. Slominska,et al.  A possible role of oxidative stress in the switch mechanism of the cell death mode from apoptosis to necrosis--studies on rho0 cells. , 2007, Mitochondrion.

[17]  M. Simon,et al.  Hypoxia-inducible factors: central regulators of the tumor phenotype. , 2007, Current opinion in genetics & development.

[18]  R. Youle,et al.  How do Bax and Bak lead to permeabilization of the outer mitochondrial membrane? , 2006, Current opinion in cell biology.

[19]  K. Tachibana,et al.  Critical role for mitochondrial oxidative phosphorylation in the activation of tumor suppressors Bax and Bak. , 2006, Journal of the National Cancer Institute.

[20]  E. Agostinelli,et al.  Non-irradiation-derived reactive oxygen species (ROS) and cancer: therapeutic implications , 2006, Amino Acids.

[21]  Yubo Sun,et al.  Oxidative phosphorylation dysfunction modulates expression of extracellular matrix--remodeling genes and invasion. , 2006, Carcinogenesis.

[22]  G. Semenza,et al.  HIF-1-mediated expression of pyruvate dehydrogenase kinase: a metabolic switch required for cellular adaptation to hypoxia. , 2006, Cell metabolism.

[23]  Koji Yoshimoto,et al.  Molecular determinants of the response of glioblastomas to EGFR kinase inhibitors. , 2005, The New England journal of medicine.

[24]  K. Ligon,et al.  Histology-Based Expression Profiling Yields Novel Prognostic Markers in Human Glioblastoma , 2005, Journal of neuropathology and experimental neurology.

[25]  R. Deberardinis,et al.  The glucose dependence of Akt-transformed cells can be reversed by pharmacologic activation of fatty acid β-oxidation , 2005, Oncogene.

[26]  C. James,et al.  Patient tumor EGFR and PDGFRA gene amplifications retained in an invasive intracranial xenograft model of glioblastoma multiforme. , 2005, Neuro-oncology.

[27]  C. Thompson,et al.  Hexokinase-mitochondria interaction mediated by Akt is required to inhibit apoptosis in the presence or absence of Bax and Bak. , 2004, Molecular cell.

[28]  R. Gillies,et al.  Why do cancers have high aerobic glycolysis? , 2004, Nature Reviews Cancer.

[29]  J. Melendez,et al.  Mitochondrial redox control of matrix metalloproteinases. , 2004, Free radical biology & medicine.

[30]  Peter Vaupel,et al.  Tumor microenvironmental physiology and its implications for radiation oncology. , 2004, Seminars in radiation oncology.

[31]  A. Alavi,et al.  Akt Stimulates Aerobic Glycolysis in Cancer Cells , 2004, Cancer Research.

[32]  V. Shoshan-Barmatz,et al.  In self-defence: hexokinase promotes voltage-dependent anion channel closure and prevents mitochondria-mediated apoptotic cell death. , 2004, The Biochemical journal.

[33]  B. Leyland-Jones,et al.  Erythropoietin to treat anaemia in patients with head and neck cancer , 2004, The Lancet.

[34]  G. Semenza Targeting HIF-1 for cancer therapy , 2003, Nature Reviews Cancer.

[35]  B. Chernyak,et al.  Oligomycin, inhibitor of the F0 part of H+-ATP-synthase, suppresses the TNF-induced apoptosis , 2002, Oncogene.

[36]  Saroj P. Mathupala,et al.  Mitochondrial bound type II hexokinase: a key player in the growth and survival of many cancers and an ideal prospect for therapeutic intervention. , 2002, Biochimica et biophysica acta.

[37]  Aftab Ahmad,et al.  Elevated expression of hexokinase II protects human lung epithelial-like A549 cells against oxidative injury. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[38]  J. Hoek,et al.  Mitochondrial Binding of Hexokinase II Inhibits Bax-induced Cytochrome c Release and Apoptosis* , 2002, The Journal of Biological Chemistry.

[39]  E. Kandel,et al.  Inhibition of early apoptotic events by Akt/PKB is dependent on the first committed step of glycolysis and mitochondrial hexokinase. , 2001, Genes & development.

[40]  S. Korsmeyer,et al.  Proapoptotic BAX and BAK: A Requisite Gateway to Mitochondrial Dysfunction and Death , 2001, Science.

[41]  F. Levi-Schaffer,et al.  Role of reactive oxygen species (ROS) in apoptosis induction , 2000, Apoptosis.

[42]  V. Mootha,et al.  tBID, a membrane-targeted death ligand, oligomerizes BAK to release cytochrome c. , 2000, Genes & development.

[43]  C. Dang,et al.  Deregulation of Glucose Transporter 1 and Glycolytic Gene Expression by c-Myc* , 2000, The Journal of Biological Chemistry.

[44]  H. Kitagawa,et al.  Calphostin C-mediated translocation and integration of Bax into mitochondria induces cytochrome c release before mitochondrial dysfunction , 2000, Cell Death and Differentiation.

[45]  Gerard I. Evan,et al.  The coordinate release of cytochrome c during apoptosis is rapid, complete and kinetically invariant , 2000, Nature Cell Biology.

[46]  M. Parliament,et al.  Modulation of oxygen consumption rate and vascular endothelial growth factor mRNA expression in human malignant glioma cells by hypoxia , 1999, British Journal of Cancer.

[47]  A. Franko,et al.  Variable presence of hypoxia in M006 human glioma spheroids and in spheroids and xenografts of clonally derived sublines. , 1998, British Journal of Cancer.

[48]  S. Korsmeyer,et al.  Regulated Targeting of BAX to Mitochondria , 1998, The Journal of cell biology.

[49]  J C Reed,et al.  Mitochondria and apoptosis. , 1998, Science.

[50]  G. Kroemer,et al.  Mitochondria as regulators of apoptosis: doubt no more. , 1998, Biochimica et biophysica acta.

[51]  C. Dang,et al.  A unique glucose-dependent apoptotic pathway induced by c-Myc. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[52]  John Calvin Reed,et al.  The Mitochondrial F0F1-ATPase proton pump is required for function of the proapoptotic protein Bax in yeast and mammalian cells. , 1998, Molecular cell.

[53]  Y. Tsujimoto,et al.  Intracellular ATP levels determine cell death fate by apoptosis or necrosis. , 1997, Cancer research.

[54]  P. Nicotera,et al.  Intracellular Adenosine Triphosphate (ATP) Concentration: A Switch in the Decision Between Apoptosis and Necrosis , 1997, The Journal of experimental medicine.

[55]  Karl Brand,et al.  Aerobic glycolysis by proliferating cells: a protective strategy against reactive oxygen species 1 , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[56]  B. Dutrillaux,et al.  Mitochondria‐bound hexokinase as target for therapy of malignant gliomas , 1995, International journal of cancer.

[57]  C. Zancanaro,et al.  Brown adipose tissue: magnetic resonance imaging and ultrastructural studies after transplantation in syngeneic rats. , 1992, Transplantation proceedings.

[58]  L. Baggetto,et al.  Deviant energetic metabolism of glycolytic cancer cells. , 1992, Biochimie.

[59]  D. Simon,et al.  Relationship of retinotopic ordering of axons in the optic pathway to the formation of visual maps in central targets , 1991, The Journal of comparative neurology.

[60]  G. Butti,et al.  Enzymes related to energy metabolism in human gliomas. , 1986, Journal of neurosurgical sciences.

[61]  O. H. Lowry,et al.  Diversity of Metabolic Patterns in Human Brain Tumors: Enzymes of Energy Metabolism and Related Metabolites and Cofactors , 1983, Journal of neurochemistry.

[62]  Lorenzo Galluzzi,et al.  Mitochondrial membrane permeabilization in cell death. , 2007, Physiological reviews.

[63]  L. B. Chen,et al.  Mitochondrial membrane potential in living cells. , 1988, Annual review of cell biology.

[64]  P. Pedersen,et al.  Tumor mitochondria and the bioenergetics of cancer cells. , 1978, Progress in experimental tumor research.