title: Laminin-332 is a substrate for hepsin, a protease associated with prostate cancer progression.
abstract: Hepsin, a cell surface protease, is widely reported to be overexpressed in more than 90% of human prostate tumors. Hepsin expression correlates with tumor progression, making it a significant marker and target for prostate cancer. Recently, it was reported that in a prostate cancer mouse model, hepsin up-regulation in tumor tissue promotes progression and metastasis. The underlying mechanisms, however, remain largely uncharacterized. Hepsin transgenic mice displayed reduced laminin-332 (Ln-332) expression in prostate tumors. This is an intriguing cue, since proteolytic processing of extracellular matrix macromolecules, such as Ln-332, is believed to be involved in cancer progression, and Ln-332 expression is lost during human prostate cancer progression. In this study, we provide the first direct evidence that hepsin cleaves Ln-332. Cleavage is specific, since it is both inhibited in a dose-dependent manner by a hepsin inhibitor (Kunitz domain-1) and does not occur when catalytically inactive hepsin is used. By Western blotting and mass spectrometry, we determined that hepsin cleaves the beta3 chain of Ln-332. N-terminal sequencing identified the cleavage site at beta3 Arg(245), in a sequence context (SQLR(245) LQGSCFC) conserved among species and in remarkable agreement with reported consensus target sequences for hepsin activity. In vitro cell migration assays showed that hepsin-cleaved Ln-332 enhanced motility of DU145 prostate cancer cells, which was inhibited by Kunitz domain-1. Further, hepsin-overexpressing LNCaP prostate cancer cells also exhibited increased migration on Ln-332. Direct cleavage of Ln-332 may be one mechanism by which hepsin promotes prostate tumor progression and metastasis, possibly by up-regulating prostate cancer cell motility.
authors.affiliation: Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, Tennessee 37232, USA.
authors.forename: Manisha
authors.initials: M
authors.lastname: Tripathi
authors.forename: Srinivas
authors.initials: S
authors.lastname: Nandana
authors.forename: Hironobu
authors.initials: H
authors.lastname: Yamashita
authors.forename: Rajkumar
authors.initials: R
authors.lastname: Ganesan
authors.forename: Daniel
authors.initials: D
authors.lastname: Kirchhofer
authors.forename: Vito
authors.initials: V
authors.lastname: Quaranta
mesh_terms: D000818:Animals; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D006801:Humans; D008297:Male; D051379:Mice; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D011471:Prostatic Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D000097794:Kalinin
chemical_list: D015815:Cell Adhesion Molecules; D009363:Neoplasm Proteins; D011480:Protease Inhibitors; D012697:Serine Endopeptidases; D000097794:Kalinin; C055645:hepsin
references.citation: J Biol Chem. 2008 Apr 11;283(15):9513-22
references.pmid: 18178964
references.citation: Int J Oncol. 2005 Feb;26(2):329-36
references.pmid: 15645116
references.citation: Bone. 2007 Dec;41(6):912-27
references.pmid: 17945547
references.citation: Invasion Metastasis. 1997;17(5):221-39
references.pmid: 9876217
references.citation: CA Cancer J Clin. 2008 Mar-Apr;58(2):71-96
references.pmid: 18287387
references.citation: Exp Dermatol. 1992 Dec;1(5):221-9
references.pmid: 1365323
references.citation: Prostate. 2001 Feb 15;46(3):240-8
references.pmid: 11170153
references.citation: Clin Exp Metastasis. 2008;25(4):377-88
references.pmid: 17972146
references.citation: Semin Cancer Biol. 1997 Feb;8(1):11-9
references.pmid: 9299577
references.citation: J Urol. 2003 Apr;169(4):1316-9
references.pmid: 12629351
references.citation: Int J Cancer. 2005 May 10;114(6):950-6
references.pmid: 15609297
references.citation: Cell Cycle. 2004 Nov;3(11):1394-7
references.pmid: 15539945
references.citation: Cancer Res. 2003 May 1;63(9):2292-9
references.pmid: 12727852
references.citation: Biochem Biophys Res Commun. 1998 Oct 9;251(1):49-55
references.pmid: 9790905
references.citation: J Mol Biol. 2005 Mar 11;346(5):1335-49
references.pmid: 15713485
references.citation: Am J Pathol. 2001 Mar;158(3):1129-35
references.pmid: 11238061
references.citation: Prostate. 2006 Sep 15;66(13):1381-90
references.pmid: 16804886
references.citation: Matrix Biol. 2004 May;23(2):75-85
references.pmid: 15246107
references.citation: Cancer Cell. 2002 Mar;1(2):203-9
references.pmid: 12086878
references.citation: Science. 1997 Jul 11;277(5323):225-8
references.pmid: 9211848
references.citation: Structure. 2003 Sep;11(9):1123-31
references.pmid: 12962630
references.citation: Bioessays. 1998 Jun;20(6):488-94
references.pmid: 9699461
references.citation: Biochem Biophys Res Commun. 2003 Apr 18;303(4):1012-7
references.pmid: 12684035
references.citation: FEBS Lett. 2005 Mar 28;579(9):1945-50
references.pmid: 15792801
references.citation: Neoplasia. 2005 Apr;7(4):380-9
references.pmid: 15967115
references.citation: Matrix Biol. 1996 Dec;15(6):369-81
references.pmid: 9049976
references.citation: Am J Pathol. 1995 Jun;146(6):1498-507
references.pmid: 7778688
references.citation: J Biol Chem. 2003 May 2;278(18):15661-8
references.pmid: 12473650
references.citation: Science. 2001 Dec 21;294(5551):2563-6
references.pmid: 11752579
references.citation: J Biol Chem. 2006 Oct 13;281(41):30439-46
references.pmid: 16908524
references.citation: J Cell Biochem. 2004 Jan 1;91(1):36-40
references.pmid: 14689579
references.citation: Clin Cancer Res. 1998 Mar;4(3):811-5
references.pmid: 9533551
references.citation: J Biol Chem. 2006 Mar 3;281(9):6020-9
references.pmid: 16365041
references.citation: Trends Cell Biol. 2003 Jul;13(7):366-75
references.pmid: 12837607
references.citation: J Invest Dermatol. 1999 Feb;112(2):177-83
references.pmid: 9989793
references.citation: J Urol. 2004 Jan;171(1):187-91
references.pmid: 14665873
references.citation: Cancer Res. 2007 May 1;67(9):4264-70
references.pmid: 17483338
references.citation: FASEB J. 2004 Feb;18(2):364-6
references.pmid: 14688206
references.citation: Am J Pathol. 2002 Jun;160(6):2169-80
references.pmid: 12057920
references.citation: Curr Opin Cell Biol. 2004 Oct;16(5):572-9
references.pmid: 15363809
references.citation: Oncogene. 2007 Mar 1;26(10):1417-27
references.pmid: 16953227
references.citation: J Cell Biol. 1991 Aug;114(3):567-76
references.pmid: 1860885
references.citation: Biochem Biophys Res Commun. 2006 May 12;343(3):692-9
references.pmid: 16563355
references.citation: J Cell Sci. 1996 Jul;109 ( Pt 7):1965-73
references.pmid: 8832419
references.citation: Cancer Cell. 2004 Aug;6(2):185-95
references.pmid: 15324701
references.citation: Biochem J. 2005 Aug 15;390(Pt 1):125-36
references.pmid: 15839837
references.citation: Nat Rev Cancer. 2007 May;7(5):370-80
references.pmid: 17457303
references.citation: Cancer Res. 2006 Apr 1;66(7):3611-9
references.pmid: 16585186
references.citation: Front Biosci. 2007 Sep 01;12:5052-9
references.pmid: 17569629
references.citation: J Biol Chem. 1995 Jan 6;270(1):66-72
references.pmid: 7814421
references.citation: Science. 2005 Mar 18;307(5716):1773-6
references.pmid: 15774758
references.citation: Trends Cell Biol. 2006 Jul;16(7):376-83
references.pmid: 16757171
references.citation: Nature. 2001 Aug 23;412(6849):822-6
references.pmid: 11518967
references.citation: Cancer Res. 2001 Jun 15;61(12):4683-8
references.pmid: 11406537
references.citation: Bioinformatics. 2004 Nov 22;20(17):3146-55
references.pmid: 15231528
references.citation: J Biol Chem. 2000 Jul 28;275(30):22728-35
references.pmid: 10806203
references.citation: Cancer Res. 2001 Aug 1;61(15):5692-6
references.pmid: 11479199
references.citation: Cancer Res. 1997 Feb 1;57(3):524-31
references.pmid: 9012485
references.citation: J Biochem. 2005 Nov;138(5):539-52
references.pmid: 16272566
references.citation: Expert Rev Anticancer Ther. 2008 Jan;8(1):33-42
references.pmid: 18095881
references.citation: Cancer Res. 2006 Dec 1;66(23):11228-37
references.pmid: 17145868
references.citation: Tumori. 2003 Jul-Aug;89(4):343-8
references.pmid: 14606634
references.citation: Nat Rev Cancer. 2002 Aug;2(8):584-93
references.pmid: 12154351
references.citation: Matrix Biol. 2005 Aug;24(5):326-32
references.pmid: 15979864
references.citation: Curr Opin Cell Biol. 1995 Oct;7(5):728-35
references.pmid: 8573349
references.citation: Cell. 2006 Nov 17;127(4):679-95
references.pmid: 17110329