Protein C and Its inhibitor in Malignancy

ABSTRACT

Activated protein C (APC) and protein C inhibitor (PCI) are the major components of the anticoagulant protein C pathway. Recently, APC and PCI have been demonstrated to play many roles not only in the regulation of hemostasis but also in cell inflammation, proliferation, apoptosis, tumor cell migration, invasion, and metastasis. Here we summarize the role of APC and PCI in malignancy.

APC increases migration of ovarian cancer cells and choriocarcinoma cells in a Transwell invasion assay in the presence of plasminogen activator inhibitor (PAI)-1; this finding suggests that APC stimulates urokinase-type plasminogen activator (uPA) by forming a complex with PAI-1 leading to activation of extracellular matrix proteases and increased invasion. It was recently reported that APC, independent of PAI-1, may increase invasion and chemotaxis of breast cancer cells by activating specific signaling pathways through endothelial protein C receptor (EPCR) and protease-activated receptor (PAR)-1. APC also increased proliferation of vascular endothelial cells and angiogenesis by EPCR-mediated activation of mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI3K), and endothelial nitric oxide synthase (eNOS) pathways.

On the other hand, we have previously reported that both uPA and PCI are synthesized in renal proximal tubular epithelial cells (RPTECs) and that PCI expression in RPTEC-derived tumor cells is significantly decreased compared with normal RPTECs. The RPTEC-derived renal carcinoma cell line Caki-1 also showed decreased expression of PCI. PCI inhibited in vitro invasive activity of Caki-1 and breast cancer cells by its protease inhibitory activity. However, PCI was found to inhibit the growth and metastatic potential of breast cancer cells independent of its protease inhibitory activity in severe combined immunodeficient mice. PCI can also inhibit angiogenesis in vivo and in vitro assays independent of its protease inhibitory activity.

Overall, these data show that APC promotes tumor cell invasion by EPCR-mediated and PAR-1-mediated protease activity and that PCI inhibits tumor cell invasion in vitro by its protease inhibitory activity and suppresses tumor cell growth, metastasis, and angiogenesis independent of its protease inhibitory activity.

KEYWORDS

Activated protein C - protein C inhibitor - SERPIN - angiogenesis

REFERENCES

• 1 Foster D, Davie E W. Characterization of a cDNA coding for human protein C.  Proc Natl Acad Sci USA. 1984;  81 4766-4770
• 2 Esmon C T, Stenflo J, Suttie J W. A new vitamin K-dependent protein. A phospholipid-binding zymogen of a serine esterase.  J Biol Chem. 1976;  251 3052-3056
• 3 Kisiel W, Ericsson L H, Davie E W. Proteolytic activation of protein C from bovine plasma.  Biochemistry. 1976;  15 4893-4900
• 4 Suzuki K, Kusumoto H, Deyashiki Y et al.. Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation.  EMBO J. 1987;  6 1891-1897
• 5 Fukudome K, Kurosawa S, Stearns-Kurosawa D J, He X, Rezaie A R, Esmon C T. The endothelial cell protein C receptor. Cell surface expression and direct ligand binding by the soluble receptor.  J Biol Chem. 1996;  271 17491-17498
• 6 Stearns-Kurosawa D J, Kurosawa S, Mollica J S, Ferrell G L, Esmon C T. The endothelial cell protein C receptor augments protein C activation by the thrombin-thrombomodulin complex.  Proc Natl Acad Sci USA. 1996;  93 10212-10216
• 7 Suzuki K, Stenflo J, Dahlbak B, Teodorsson B. Inactivation of human coagulation factor V by activated protein C.  J Biol Chem. 1983;  258 1914-1920
• 8 Marlar R A, Kleiss A J, Griffin J H. Human protein C: inactivation of factors V and VIII in plasma by the activated molecule.  Ann N Y Acad Sci. 1981;  370 303-310
• 9 Walker F J. Regulation of activated protein C by a new protein. A possible function for bovine protein S.  J Biol Chem. 1980;  255 5521-5524
• 10 Walker F J. Regulation of activated protein C by protein S. The role of phospholipid in factor Va inactivation.  J Biol Chem. 1981;  256 11128-11131
• 11 Hoffman M, Monroe III D M. A cell-based model of hemostasis.  Thromb Haemost. 2001;  85 958-965
• 12 Esmon C T. The protein C pathway.  Chest. 2003;  124 26S-32S
• 13 Branson H E, Katz J, Marble R, Griffin J H. Inherited protein C deficiency and coumarin-responsive chronic relapsing purpura fulminans in a newborn infant.  Lancet. 1983;  2 1165-1168
• 14 Schwarz H P, Fischer M, Hopmeier P, Batard M A, Griffin J H. Plasma protein S deficiency in familial thrombotic disease.  Blood. 1984;  64 1297-1300
• 15 Ohlin A K, Marlar R A. The first mutation identified in the thrombomodulin gene in a 45-year-old man presenting with thromboembolic disease.  Blood. 1995;  85 330-336
• 16 Gu J M, Crawley J T, Ferrell G et al.. Disruption of the endothelial cell protein C receptor gene in mice causes placental thrombosis and early embryonic lethality.  J Biol Chem. 2002;  277 43335-43343
• 17 Taylor Jr F B, Stearns-Kurosawa D J, Kurosawa S et al.. The endothelial cell protein C receptor aids in host defense against Escherichia coli sepsis.  Blood. 2000;  95 1680-1686
• 18 Bernard G R, Vincent J L, Laterre P F et al.. Efficacy and safety of recombinant human activated protein C for severe sepsis.  N Engl J Med. 2001;  344 699-709
• 19 Yuda H, Adachi Y, Taguchi O et al.. Activated protein C inhibits bronchial hyperresponsiveness and Th2 cytokine expression in mice.  Blood. 2004;  103 2196-2204
• 20 Joyce D E, Gelbert L, Ciaccia A, DeHoff B, Grinnell B W. Gene expression profile of antithrombotic protein c defines new mechanisms modulating inflammation and apoptosis.  J Biol Chem. 2001;  276 11199-11203
• 21 Cheng T, Liu D, Griffin J H et al.. Activated protein C blocks p53-mediated apoptosis in ischemic human brain endothelium and is neuroprotective.  Nat Med. 2003;  9 338-342
• 22 Feistritzer C, Sturn D H, Kaneider N C, Djanani A, Wiedermann C J. Endothelial protein C receptor-dependent inhibition of human eosinophil chemotaxis by protein C.  J Allergy Clin Immunol. 2003;  112 375-381
• 23 Feistritzer C, Mosheimer B A, Sturn D H, Riewald M, Patsch J R, Wiedermann C J. Endothelial protein C receptor-dependent inhibition of migration of human lymphocytes by protein C involves epidermal growth factor receptor.  J Immunol. 2006;  176 1019-1025
• 24 Jackson C J, Xue M, Thompson P et al.. Activated protein C prevents inflammation yet stimulates angiogenesis to promote cutaneous wound healing.  Wound Repair Regen. 2005;  13 284-294
• 25 Suzuki K, Nishioka J, Hashimoto S. Protein C inhibitor. Purification from human plasma and characterization.  J Biol Chem. 1983;  258 163-168
• 26 Suzuki K, Deyashiki Y, Nishioka J, Akira M, Yamamoto S, Hashimoto S. Characterization of a cDNA for human protein C inhibitor: A new member of the plasma serine protease inhibitor superfamily.  J Biol Chem. 1987;  262 611-616
• 27 Suzuki K, Nishioka J, Kusumoto H, Hashimoto S. Mechanism of inhibition of activated protein C by protein C inhibitor.  J Biochem (Tokyo). 1984;  85 187-195
• 28 Rezaie A R, Cooper S T, Church F C, Esmon C T. Protein C inhibitor is a potent inhibitor of the thrombin-thrombomodulin complex.  J Biol Chem. 1995;  270 25336-25339
• 29 Meijers J CM, Kanters D H, Vlooswijk R A, van Erp H E, Hessing M, Bouma B N. Inactivation of human plasma kallikrein and factor XIa by protein C inhibitor.  Biochemistry. 1988;  27 4231-4237
• 30 Geiger M, Huber K, Wojta J et al.. Complex formation between urokinase and plasma protein C inhibitor in vitro and in vivo.  Blood. 1989;  74 722-728
• 31 Suzuki K. Activated protein C inhibitor.  Semin Thromb Hemost. 1984;  10 154-161
• 32 Stief T W, Radtke K P, Heimburger N. Inhibition of urokinase by protein C-inhibitor (PCI). Evidence for identity of PCI and plasminogen activator inhibitor 3.  Biol Chem Hoppe Seyler. 1987;  368 1427-1433
• 33 Hermans J M, Jones R, Stone R. Rapid inhibition of the sperm protease acrosin by protein C inhibitor.  Biochemistry. 1994;  33 5440-5444
• 34 Christensson A, Lilja H. Complex formation between protein C inhibitor and prostate-specific antigen in vitro and human semen.  Eur J Biochem. 1994;  220 45-53
• 35 Hayashi T, Nishioka J, Nakagawa N et al.. Protein C inhibitor directly and potently inhibits activated hepatocyte growth factor activator.  J Thromb Haemost. 2007;  5 1477-1485
• 36 Andreasen P A, Egelund R, Petersen H H. The plasminogen activation system in tumor growth, invasion, and metastasis.  Cell Mol Life Sci. 2000;  57 25-40
• 37 Andreasen P A, Kjoller L, Christensen L, Duffy M J. The urokinase-type plasminogen activator system in cancer metastasis.  Int J Cancer. 1997;  72 1-22
• 38 Sieben N L, Oosting J, Flanagan A M et al.. Differential gene expression in ovarian tumors reveals Dusp 4 and Serpina 5 as key regulators for benign behavior of serous borderline tumors.  J Clin Oncol. 2005;  23 7257-7264
• 39 Cao Y, Becker C, Lundwall A et al.. Expression of protein C inhibitor (PCI) in benign and malignant prostate tissues.  Prostate. 2003;  57 196-204
• 40 Wakita T, Hayashi T, Nishioka J et al.. Regulation of carcinoma cell invasion by protein C inhibitor whose expression is decreased in renal cell carcinoma.  Int J Cancer. 2004;  108 516-523
• 41 Glasscock L N, Rehault S M, Gregory C W et al.. Protein C inhibitor (plasminogen activator inhibitor-3) expression in the CWR22 prostate cancer xenograft.  Exp Mol Pathol. 2005;  79 23-32
• 42 Palmieri D, Lee J W, Juliano R L, Church F C. Plasminogen activator inhibitor-1 and -3 increase cell adhesion and motility of MDA-MB-435 breast cancer cells.  J Biol Chem. 2002;  277 40950-40957
• 43 McMahon G A, Petitclerc E, Stefansson S et al.. Plasminogen activator inhibitor-1 regulates tumor growth and angiogenesis.  J Biol Chem. 2001;  276 33964-33968
• 44 O'Reilly M S, Pirie-Shepherd S, Lane W S, Folkman J. Antiangiogenic activity of the cleaved conformation of the serpin antithrombin.  Science. 1999;  285 1926-1928
• 45 Huang H, Campbell S C, Nelius T et al.. Alpha1-antitrypsin inhibits angiogenesis and tumor growth.  Int J Cancer. 2004;  112 1042-1048
• 46 Kobayashi H, Moniwa N, Gotoh J, Sugimura M, Terao T. Role of activated protein C in facilitating basement membrane invasion by tumor cells.  Cancer Res. 1994;  54 261-267
• 47 Beaulieu A, Church F C. Activated protein C promotes breast cancer cell migration through interactions with EPCR and PAR-1.  Exp Cell Res. 2007;  313 677-687
• 48 Uchiba M, Okajima K, Oike Y et al.. Activated protein C induces endothelial cell proliferation by mitogen-activated protein kinase activation in vitro and angiogenesis in vivo.  Circ Res. 2004;  95 34-41
• 49 Asanuma K, Yoshikawa T, Hayashi T et al.. Protein C inhibitor inhibits breast cancer cell growth, metastasis and angiogenesis independently of its protease inhibitory activity.  Int J Cancer. 2007;  121 955-965
• 50 Folkman J, Shing Y. Control of angiogenesis by heparin and other sulfated polysaccharides.  Adv Exp Med Biol. 1992;  313 355-364
• 51 Miao R Q, Chen V, Chao L, Chao J. Structural elements of kallistatin required for inhibition of angiogenesis.  Am J Physiol Cell Physiol. 2003;  284 C1604-C1613
• 52 Zhang W, Chuang Y J, Swanson R et al.. Antiangiogenic antithrombin down-regulates the expression of the proangiogenic heparan sulfate proteoglycan, perlecan, in endothelial cells.  Blood. 2004;  103 1185-1191