title: NADPH oxidase NOX4 supports renal tumorigenesis by promoting the expression and nuclear accumulation of HIF2α.
abstract: Most sporadically occurring renal tumors include a functional loss of the tumor suppressor von Hippel Lindau (VHL). Development of VHL-deficient renal cell carcinoma (RCC) relies upon activation of the hypoxia-inducible factor-2α (HIF2α), a master transcriptional regulator of genes that drive diverse processes, including angiogenesis, proliferation, and anaerobic metabolism. In determining the critical functions for HIF2α expression in RCC cells, the NADPH oxidase NOX4 has been identified, but the pathogenic contributions of NOX4 to RCC have not been evaluated directly. Here, we report that NOX4 silencing in VHL-deficient RCC cells abrogates cell branching, invasion, colony formation, and growth in a murine xenograft model RCC. These alterations were phenocopied by treatment of the superoxide scavenger, TEMPOL, or by overexpression of manganese superoxide dismutase or catalase. Notably, NOX4 silencing or superoxide scavenging was sufficient to block nuclear accumulation of HIF2α in RCC cells. Our results offer direct evidence that NOX4 is critical for renal tumorigenesis and they show how NOX4 suppression and VHL re-expression in VHL-deficient RCC cells are genetically synonymous, supporting development of therapeutic regimens aimed at NOX4 blockade.
authors.affiliation: University of Pittsburgh, Department of Urology and University of Pittsburgh Cancer Institute.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Gregg
authors.affiliation: University of Pittsburgh, Department of Urology and University of Pittsburgh Cancer Institute.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Turner
authors.affiliation: University of Pittsburgh, Department of Urology and University of Pittsburgh Cancer Institute.
authors.forename: Guimin
authors.initials: G
authors.lastname: Chang
authors.affiliation: University of Pittsburgh, Department of Urology and University of Pittsburgh Cancer Institute.
authors.forename: Disha
authors.initials: D
authors.lastname: Joshi
authors.affiliation: University of Massachusetts, Department of Surgery.
authors.forename: Ye
authors.initials: Y
authors.lastname: Zhan
authors.affiliation: University of Pittsburgh, Department of Urology and University of Pittsburgh Cancer Institute.
authors.forename: Li
authors.initials: L
authors.lastname: Chen
authors.affiliation: University of Pittsburgh, Department of Urology and University of Pittsburgh Cancer Institute.
authors.forename: Jodi K
authors.initials: JK
authors.lastname: Maranchie
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000096:biosynthesis / Q000378:metabolism*; D002292:Carcinoma, Renal Cell / Q000473:pathology*; D002374:Catalase / Q000096:biosynthesis; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000473:pathology; D003497:Cyclic N-Oxides / Q000494:pharmacology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007668:Kidney / Q000473:pathology; D007680:Kidney Neoplasms / Q000235:genetics / Q000473:pathology*; D051379:Mice; D016513:Mice, SCID; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000235:genetics*; D009368:Neoplasm Transplantation; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D034622:RNA Interference; D013113:Spin Labels; D013482:Superoxide Dismutase / Q000096:biosynthesis; D013481:Superoxides / Q000378:metabolism; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics / Q000378:metabolism*; D000099162:Endothelial PAS Domain-Containing Protein 1
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D002374:Catalase; D003497:Cyclic N-Oxides; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; D011500:Protein Synthesis Inhibitors; D013113:Spin Labels; D013482:Superoxide Dismutase; D013481:Superoxides; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D000099162:Endothelial PAS Domain-Containing Protein 1; C498910:NOX4 protein, human; C497446:VHL protein, human; C001803:tempol
references.citation: Cancer Res. 2002 May 15;62(10):2957-61
references.pmid: 12019178
references.citation: Antioxid Redox Signal. 2005 Mar-Apr;7(3-4):456-64
references.pmid: 15706093
references.citation: Cancer. 2009 May 15;115(10 Suppl):2298-305
references.pmid: 19402060
references.citation: Free Radic Biol Med. 2004 May 15;36(10):1279-88
references.pmid: 15110393
references.citation: Mol Cell Biol. 2003 Dec;23(24):9361-74
references.pmid: 14645546
references.citation: Am J Physiol Heart Circ Physiol. 2007 Oct;293(4):H2085-92
references.pmid: 17644566
references.citation: Cancer Cell. 2002 Apr;1(3):247-55
references.pmid: 12086861
references.citation: Cell Signal. 2006 Jan;18(1):69-82
references.pmid: 15927447
references.citation: Mol Cell Biol. 1999 Sep;19(9):5902-12
references.pmid: 10454537
references.citation: Cancer Cell. 2002 Apr;1(3):237-46
references.pmid: 12086860
references.citation: Am J Pathol. 2010 May;176(5):2447-55
references.pmid: 20304964
references.citation: PLoS Biol. 2003 Dec;1(3):E83
references.pmid: 14691554
references.citation: J Cell Biochem. 2003 Oct 1;90(2):304-14
references.pmid: 14505347
references.citation: Cancer Res. 2005 Oct 15;65(20):9190-3
references.pmid: 16230378
references.citation: Biochem Biophys Res Commun. 2009 Feb 6;379(2):445-50
references.pmid: 19116140
references.citation: Curr Opin Oncol. 2011 May;23(3):283-9
references.pmid: 21330923
references.citation: Neurobiol Dis. 2010 Jan;37(1):118-29
references.pmid: 19804834
references.citation: Arterioscler Thromb Vasc Biol. 2004 Apr;24(4):677-83
references.pmid: 14670934
references.citation: J Biol Chem. 2004 Aug 13;279(33):34643-54
references.pmid: 15155719
references.citation: Hypertension. 2008 Jul;52(1):143-9
references.pmid: 18474828
references.citation: J Biol Chem. 2000 Aug 18;275(33):25130-8
references.pmid: 10833514
references.citation: PLoS One. 2012;7(1):e30712
references.pmid: 22303451
references.citation: Cancer Res. 2009 Mar 15;69(6):2375-83
references.pmid: 19276362
references.citation: Antioxid Redox Signal. 2009 Apr;11(4):747-64
references.pmid: 18783311
references.citation: Oncogene. 2005 Dec 8;24(55):8154-66
references.pmid: 16170370
references.citation: Antioxid Redox Signal. 2010 Aug 15;13(4):425-36
references.pmid: 20039838
references.citation: Free Radic Biol Med. 2009 Nov 1;47(9):1239-53
references.pmid: 19628035
references.citation: Cardiovasc Res. 2008 Nov 1;80(2):299-308
references.pmid: 18567639
references.citation: Cancer Res. 2009 Mar 15;69(6):2647-54
references.pmid: 19276355
references.citation: Free Radic Biol Med. 2012 Oct 1;53(7):1489-99
references.pmid: 22728268
references.citation: Exp Mol Med. 2010 Apr 30;42(4):270-9
references.pmid: 20177149
references.citation: Nat Med. 1995 Aug;1(8):822-6
references.pmid: 7585187
references.citation: Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):8010-4
references.pmid: 10869423
references.citation: Cancer Discov. 2011 Aug;1(3):222-35
references.pmid: 22037472
references.citation: Mol Cell Biol. 2005 Jul;25(13):5675-86
references.pmid: 15964822
references.citation: Sci Signal. 2009 Sep 15;2(88):ra53
references.pmid: 19755709
references.citation: Cancer Res. 2000 Aug 15;60(16):4419-25
references.pmid: 10969787
references.citation: J Biol Chem. 2006 Aug 11;281(32):22575-85
references.pmid: 16760477
references.citation: J Cell Biochem. 2008 Apr 15;103(6):1989-98
references.pmid: 17973296
references.citation: CA Cancer J Clin. 2013 Jan;63(1):11-30
references.pmid: 23335087
references.citation: J Biol Chem. 2007 Mar 16;282(11):8019-26
references.pmid: 17200123
references.citation: Cancer Res. 2008 Apr 15;68(8):2781-8
references.pmid: 18413745
references.citation: Vascul Pharmacol. 2012 May-Jun;56(5-6):245-51
references.pmid: 22366374
references.citation: Am J Physiol. 1994 Dec;267(6 Pt 1):L815-22
references.pmid: 7810685