title: Nitroxoline impairs tumor progression in vitro and in vivo by regulating cathepsin B activity.
abstract: Cathepsin B is a ubiquitously expressed lysosomal cysteine protease that participates in protein turnover within lysosomes. However, its protein and activity levels have been shown to be increased in cancer. Cathepsin B endopeptidase activity is involved in the degradation of extracellular matrix, a process that promotes tumor invasion, metastasis and angiogenesis. Previously, we reported an established antibiotic nitroxoline as a potent and selective inhibitor of cathepsin B. In the present study, we elucidated its anti-tumor properties in in vitro and in vivo tumor models. Tumor and endothelial cell lines with high levels of active cathepsin B were selected for functional analysis of nitroxoline in vitro. Nitroxoline significantly reduced extracellular DQ-collagen IV degradation by all evaluated cancer cell lines using spectrofluorimetry. Nitroxoline also markedly decreased tumor cell invasion monitored in real time and reduced the invasive growth of multicellular tumor spheroids, used as a 3D in vitro model of tumor invasion. Additionally, endothelial tube formation was significantly reduced by nitroxoline in an in vitro angiogenesis assay. Finally, nitroxoline significantly abrogated tumor growth, angiogenesis and metastasis in vivo in LPB fibrosarcoma and MMTV-PyMT breast cancer mouse models. Overall, our results designate nitroxoline as a promising drug candidate for anti-cancer treatment.
authors.affiliation: Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia.
authors.forename: Bojana
authors.initials: B
authors.lastname: Mirković
authors.affiliation: Department of Experimental Oncology, Institute of Oncology Ljubljana, Ljubljana, Slovenia.
authors.forename: Boštjan
authors.initials: B
authors.lastname: Markelc
authors.affiliation: Department of Biochemistry and Molecular and Structural Biology, Jožef Stefan Institute, Ljubljana, Slovenia.
authors.forename: Miha
authors.initials: M
authors.lastname: Butinar
authors.affiliation: Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia.
authors.forename: Ana
authors.initials: A
authors.lastname: Mitrović
authors.affiliation: Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia.
authors.forename: Izidor
authors.initials: I
authors.lastname: Sosič
authors.affiliation: Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia.
authors.forename: Stanislav
authors.initials: S
authors.lastname: Gobec
authors.affiliation: Department of Biochemistry and Molecular and Structural Biology, Jožef Stefan Institute, Ljubljana, Slovenia.
authors.forename: Olga
authors.initials: O
authors.lastname: Vasiljeva
authors.affiliation: Department of Biochemistry and Molecular and Structural Biology, Jožef Stefan Institute, Ljubljana, Slovenia.|Centre of Excellence for Integrated Approaches in Chemistry and Biology of Proteins, Ljubljana, Slovenia.|Faculty of Chemistry and Chemical Technology, University of Ljubljana, Ljubljana, Slovenia.
authors.forename: Boris
authors.initials: B
authors.lastname: Turk
authors.affiliation: Department of Experimental Oncology, Institute of Oncology Ljubljana, Ljubljana, Slovenia.
authors.forename: Maja
authors.initials: M
authors.lastname: Čemažar
authors.affiliation: Department of Experimental Oncology, Institute of Oncology Ljubljana, Ljubljana, Slovenia.
authors.forename: Gregor
authors.initials: G
authors.lastname: Serša
authors.affiliation: Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia.|Department of Biotechnology, Jožef Stefan Institute, Ljubljana, Slovenia.
authors.forename: Janko
authors.initials: J
authors.lastname: Kos
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000494:pharmacology*; D000818:Animals; D016022:Case-Control Studies; D002401:Cathepsin B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D048708:Cell Growth Processes / Q000187:drug effects; D045744:Cell Line, Tumor; D018450:Disease Progression; D042783:Endothelial Cells / Q000187:drug effects; D005260:Female; D005354:Fibrosarcoma / Q000098:blood supply / Q000188:drug therapy / Q000201:enzymology; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009362:Neoplasm Metastasis; D009600:Nitroquinolines / Q000494:pharmacology*; D011897:Random Allocation; D012513:Sarcoma, Experimental / Q000098:blood supply / Q000188:drug therapy / Q000201:enzymology; D018874:Spheroids, Cellular
chemical_list: D043925:Angiogenesis Inducing Agents; D009600:Nitroquinolines; C005308:nitroxoline; D002401:Cathepsin B
keywords: angiogenesis; cathepsin B; metastasis; nitroxoline; tumor invasion
references.citation: Cancer Metastasis Rev. 2003 Jun-Sep;22(2-3):271-86
references.pmid: 12785001
references.citation: Exp Cell Res. 2003 Feb 15;283(2):206-14
references.pmid: 12581740
references.citation: Int J Cancer. 1972 May 15;9(3):461-9
references.pmid: 4660932
references.citation: Mol Carcinog. 1991;4(1):25-35
references.pmid: 2009132
references.citation: FEBS Lett. 1991 Mar 25;280(2):311-5
references.pmid: 2013329
references.citation: EMBO J. 1991 Sep;10(9):2321-30
references.pmid: 1868826
references.citation: Biochem J. 1992 Feb 15;282 ( Pt 1):273-8
references.pmid: 1540143
references.citation: Mol Cell Biol. 1992 Mar;12(3):954-61
references.pmid: 1312220
references.citation: J Cell Sci. 1994 Feb;107 ( Pt 2):373-84
references.pmid: 8207069
references.citation: J Biol Chem. 1997 Jan 10;272(2):1197-202
references.pmid: 8995421
references.citation: Biochemistry. 1997 Oct 14;36(41):12608-15
references.pmid: 9376367
references.citation: Oncol Rep. 1998 Nov-Dec;5(6):1349-61
references.pmid: 9769367
references.citation: Clin Cancer Res. 1997 Oct;3(10):1815-22
references.pmid: 9815568
references.citation: Clin Cancer Res. 1999 Mar;5(3):559-67
references.pmid: 10100707
references.citation: Eur J Biochem. 1999 Aug;264(1):100-9
references.pmid: 10447678
references.citation: IEEE Trans Med Imaging. 2005 Apr;24(4):549-53
references.pmid: 15822812
references.citation: J Med Chem. 2005 Dec 15;48(25):8045-54
references.pmid: 16335928
references.citation: Genes Dev. 2006 Mar 1;20(5):543-56
references.pmid: 16481467
references.citation: J Cell Biochem. 2006 Apr 15;97(6):1230-40
references.pmid: 16315320
references.citation: Cancer Res. 2006 May 15;66(10):5242-50
references.pmid: 16707449
references.citation: Curr Med Chem. 2006;13(19):2309-27
references.pmid: 16918357
references.citation: Nat Rev Drug Discov. 2006 Sep;5(9):785-99
references.pmid: 16955069
references.citation: Nat Rev Cancer. 2006 Oct;6(10):764-75
references.pmid: 16990854
references.citation: J Biomol Screen. 2006 Dec;11(8):922-32
references.pmid: 16973921
references.citation: Curr Pharm Des. 2007;13(4):387-403
references.pmid: 17311556
references.citation: Methods Mol Med. 2007;140:141-51
references.pmid: 18085207
references.citation: Oncogene. 2008 Jul 10;27(30):4191-9
references.pmid: 18345026
references.citation: Cancer Res. 2008 Nov 15;68(22):9147-56
references.pmid: 19010886
references.citation: Exp Cell Res. 2009 Apr 15;315(7):1234-46
references.pmid: 19331819
references.citation: Cancer Res. 2009 May 15;69(10):4537-44
references.pmid: 19435903
references.citation: FEBS J. 2009 Sep;276(17):4739-51
references.pmid: 19656187
references.citation: Pathol Oncol Res. 2009 Dec;15(4):711-23
references.pmid: 19434518
references.citation: Proc Natl Acad Sci U S A. 2010 Feb 9;107(6):2497-502
references.pmid: 20133781
references.citation: Curr Top Med Chem. 2010;10(7):696-707
references.pmid: 20337581
references.citation: Neoplasia. 2010 Jun;12(6):485-98
references.pmid: 20563251
references.citation: Mol Cancer. 2010;9:207
references.pmid: 20684763
references.citation: J Natl Cancer Inst. 2010 Dec 15;102(24):1855-73
references.pmid: 21088277
references.citation: Biotechnol Bioeng. 2003 Jul 20;83(2):173-80
references.pmid: 12768623
references.citation: Oncogene. 2011 Jan 6;30(1):54-64
references.pmid: 20818432
references.citation: ChemMedChem. 2011 Aug 1;6(8):1351-6
references.pmid: 21598397
references.citation: Nat Nanotechnol. 2011 Sep;6(9):594-602
references.pmid: 21822252
references.citation: Biochim Biophys Acta. 2012 Jan;1824(1):68-88
references.pmid: 22024571
references.citation: Nat Commun. 2012;3:751
references.pmid: 22434194
references.citation: Breast Cancer Res. 2011;13(6):R115
references.pmid: 22093547
references.citation: Eur J Immunol. 2012 Dec;42(12):3429-41
references.pmid: 23018451
references.citation: Neuro Oncol. 2015 Jan;17(1):53-62
references.pmid: 25074541
references.citation: Int J Dev Neurosci. 1999 Aug-Oct;17(5-6):483-94
references.pmid: 10571410
references.citation: Chem Biol. 1999 Nov;6(11):823-33
references.pmid: 10574784
references.citation: Clin Chim Acta. 2000 Feb 15;291(2):113-35
references.pmid: 10675719
references.citation: Crit Rev Oncol Hematol. 2000 Nov-Dec;36(2-3):123-39
references.pmid: 11033302
references.citation: J Biol Chem. 2001 Jan 12;276(2):944-51
references.pmid: 11016923
references.citation: Neoplasia. 2000 Nov-Dec;2(6):496-504
references.pmid: 11228542
references.citation: Biol Chem. 2001 May;382(5):853-7
references.pmid: 11517941
references.citation: Clin Chim Acta. 2004 Apr;342(1-2):41-69
references.pmid: 15026265