Cathepsin L Induces Proangiogenic Changes in Human Omental Microvascular Endothelial Cells via Activation of the ERK1/2 Pathway

Author(s): Md Zahidul I. Pranjol, Nicholas J. Gutowski, Michael Hannemann, Jacqueline L. Whatmore*.

Journal Name: Current Cancer Drug Targets

Volume 19 , Issue 3 , 2019

  Journal Home
Translate in Chinese
Submit Manuscript
Submit Proposal

Graphical Abstract:


Background: Metastasis still remains the major cause of therapeutic failure, poor prognosis and high mortality in epithelial ovarian cancer (EOC) patients. Previously, we showed that EOC cells secrete a range of factors with potential pro-angiogenic activity, in disease-relevant human omental microvascular endothelial cells (HOMECs), including the lysosomal protease cathepsin L (CathL). Thus, the aim of this study was to examine potential pro-proliferative and pro-migratory effects of CathL in HOMECs and the activated signalling pathways, and whether these proangiogenic responses are dependent on CathL-catalytic activity.

Methods: HOMECs proliferation was investigated using WST-1, BrdU and CyQUANT assays. Cell migration was examined using a Cultrex Cell 96 transwell migration assay. Enzyme activity was assayed at various pHs using the CathL-specific fluorogenic substrate FY-CHO. Activation of cell signalling pathways was tested using a commercially available phosphokinase array and intact cellbased ELISAs.

Results: We showed for the first time that CathL has a potent pro-proliferative and pro-migratory effect on HOMECs. For instance, CathL significantly increases HOMEC proliferation (134.8±14.7% vs control 100%) and migration (146.6±17.3% vs control 100%). Our data strongly suggest that these proangiogenic effects of CathL are mediated via a non-proteolytic mechanism. Finally, we show that CathL-induced activation of the ERK1/2 pathway is involved in inducing these cellular effects in HOMECs.

Conclusion: These data suggest that CathL acts as an extracellular ligand and plays an important pro-angiogenic, and thus pro-metastatic, role during EOC metastasis to the omentum, by activating the omental microvasculature, and thus can potentially be targeted therapeutically in the future.

Keywords: Cathepsin L, non-proteolytic, proliferation, migration, angiogenesis, metastasis.

Lengyel E. Ovarian cancer development and metastasis. Am J Pathol 2010; 177(3): 1053-64.
van der Bilt AR, van der Zee AG, de Vries EG, et al. Multiple VEGF family members are simultaneously expressed in ovarian cancer: a proposed model for bevacizumab resistance. Curr Pharm Des 2012; 18(25): 3784-92.
Lin Z, Liu Y, Sun Y, He X. Expression of Ets-1, Ang-2 and maspin in ovarian cancer and their role in tumor angiogenesis. J Exp Clin Cancer Res 2011; 30: 31.
Tanaka Y, Miyamoto S, Suzuki SO, et al. Clinical significance of heparin-binding epidermal growth factor-like growth factor and a disintegrin and metalloprotease 17 expression in human ovarian cancer. Clin Cancer Res 2005; 11(13): 4783-92.
Zhong H, De Marzo AM, Laughner E, et al. Overexpression of hypoxia-inducible factor 1alpha in common human cancers and their metastases. Cancer Res 1999; 59(22): 5830-5.
Toutirais O, Chartier P, Dubois D, et al. Constitutive expression of TGF-beta1, interleukin-6 and interleukin-8 by tumor cells as a major component of immune escape in human ovarian carcinoma. Eur Cytokine Netw 2003; 14(4): 246-55.
Hollingsworth HC, Kohn EC, Steinberg SM, Rothenberg ML, Merino MJ. Tumor angiogenesis in advanced stage ovarian carcinoma. Am J Pathol 1995; 147(1): 33-41.
Abulafia O, Triest WE, Sherer DM. Angiogenesis in primary and metastatic epithelial ovarian carcinoma. Am J Obstet Gynecol 1997; 177(3): 541-7.
Winiarski BK, Wolanska KI, Rai S, et al. Epithelial ovarian cancer-induced angiogenic phenotype of human omental microvascular endothelial cells may occur independently of VEGF signaling. Transl Oncol 2013; 6(6): 703-14.
Teoh D, Secord AA. Antiangiogenic agents in combination with chemotherapy for the treatment of epithelial ovarian cancer. Int J Gynecol Cancer 2012; 22(3): 348-59.
Kirschke H, Langner J, Wiederanders B, Ansorge S, Bohley P. Cathepsin, L. A new proteinase from rat-liver lysosomes. Eur J Biochem 1977; 74(2): 293-301.
Kominami E, Ueno T, Muno D, Katunuma N. The selective role of cathepsins B and D in the lysosomal degradation of endogenous and exogenous proteins. FEBS Lett 1991; 287(1-2): 189-92.
Cuvier C, Jang A, Hill RP. Exposure to hypoxia, glucose starvation and acidosis: effect on invasive capacity of murine tumor cells and correlation with cathepsin (L + B) secretion. Clin Exp Metastasis 1997; 15(1): 19-25.
Zhang W, Wang S, Wang Q, Yang Z, Pan Z, Li L. Overexpression of cysteine cathepsin L is a marker of invasion and metastasis in ovarian cancer. Oncol Rep 2014; 31(3): 1334-42.
Winiarski BK, Cope N, Alexander M, et al. Clinical relevance of increased endothelial and mesothelial expression of proangiogenic proteases and vegfa in the omentum of patients with metastatic ovarian high-grade serous carcinoma. Transl Oncol 2014; 7(2): 267-76.
Winiarski BK, Acheson N, Gutowski NJ, McHarg S, Whatmore JL. An improved and reliable method for isolation of microvascular endothelial cells from human omentum. Microcirculation 2011; 18(8): 635-45.
Pranjol MZI, Gutowski NJ, Hannemann M, Whatmore JL. Cathepsin D non-proteolytically induces proliferation and migration in human omental microvascular endothelial cells via activation of the ERK1/2 and PI3K/AKT pathways. Biochim Biophys Acta 2017; 1865(1): 25-33.
Bird TA, Schule HD, Delaney PB, Sims JE, Thoma B, Dower SK. Evidence that MAP (mitogen-activated protein) kinase activation may be a necessary but not sufficient signal for a restricted subset of responses in IL-1-treated epidermoid cells. Cytokine 1992; 4(6): 429-40.
Konopatskaya O, Shore AC, Tooke JE, Whatmore JL. A role for heterotrimeric GTP-binding proteins and ERK1/2 in insulin-mediated, nitric-oxide-dependent, cyclic GMP production in human umbilical vein endothelial cells. Diabetologia 2005; 48(3): 595-604.
Pranjol MZ, Gutowski N, Hannemann M, Whatmore J. The potential role of the proteases cathepsin D and Cathepsin L in the progression and metastasis of epithelial ovarian cancer. Biomolecules 2015; 5(4): 3260-79.
Weitoft T, Larsson A, Manivel VA, Lysholm J, Knight A, Ronnelid J. Cathepsin S and cathepsin L in serum and synovial fluid in rheumatoid arthritis with and without autoantibodies. Rheumatology 2015; 54(10): 1923-8.
Fiebiger E, Maehr R, Villadangos J, et al. Invariant chain controls the activity of extracellular cathepsin L. J Exp Med 2002; 196(9): 1263-9.
Kominami E, Tsukahara T, Hara K, Katunuma N. Biosyntheses and processing of lysosomal cysteine proteinases in rat macrophages. FEBS Lett 1988; 231(1): 225-8.
Nishimura Y, Furuno K, Kato K. Biosynthesis and processing of lysosomal cathepsin L in primary cultures of rat hepatocytes. Arch Biochem Biophys 1988; 263(1): 107-16.
Dong JM, Sahagian GG. Basis for low affinity binding of a lysosomal cysteine protease to the cation-independent mannose 6-phosphate receptor. J Boil Chem 1990; 265(8): 4210-7.
Lang L, Reitman M, Tang J, Roberts RM, Kornfeld S. Lysosomal enzyme phosphorylation. Recognition of a protein-dependent determinant allows specific phosphorylation of oligosaccharides present on lysosomal enzymes. J Boil Chem 1984; 259(23): 14663-71.
Kornfeld S. Trafficking of lysosomal enzymes in normal and disease states. J Clin Invest 1986; 77(1): 1-6.
Samie MA, Xu H. Lysosomal exocytosis and lipid storage disorders. J Lipid Res 2014; 55(6): 1995-009.
Mason RW, Gal S, Gottesman MM. The identification of the major excreted protein (MEP) from a transformed mouse fibroblast cell line as a catalytically active precursor form of cathepsin L. Biochem J 1987; 248(2): 449-54.
von Figura K, Hasilik A. Lysosomal enzymes and their receptors. Annu Rev Biochem 1986; 55: 167-93.
Dahms NM, Lobel P, Kornfeld S. Mannose 6-phosphate receptors and lysosomal enzyme targeting. J Boil Chem 1989; 264(21): 12115-8.
Dong JM, Prence EM, Sahagian GG. Mechanism for selective secretion of a lysosomal protease by transformed mouse fibroblasts. J Boil Chem 1989; 264(13): 7377-83.
Maciewicz RA, Wotton SF. Degradation of cartilage matrix components by the cysteine proteinases, cathepsins B and L. Biomed Biochim Acta 1991; 50(4-6): 561-4.
Maciewicz RA, Wotton SF, Etherington DJ, Duance VC. Susceptibility of the cartilage collagens types II, IX and XI to degradation by the cysteine proteinases, cathepsins B and L. FEBS Lett 1990; 269(1): 189-93.
Nguyen Q, Mort JS, Roughley PJ. Cartilage proteoglycan aggregate is degraded more extensively by cathepsin L than by cathepsin B. Biochem J 1990; 266(2): 569-73.
Nosaka AY, Kanaori K, Teno N, et al. Conformational studies on the specific cleavage site of Type I collagen (alpha-1) fragment (157-192) by cathepsins K and L by proton NMR spectroscopy. Bioorg Med Chem 1999; 7(2): 375-9.
Mason RW, Johnson DA, Barrett AJ, Chapman HA. Elastinolytic activity of human cathepsin L. Biochem J 1986; 233(3): 925-7.
Ishidoh K, Kominami E. Procathepsin L degrades extracellular matrix proteins in the presence of glycosaminoglycans in vitro. Biochem Biophys Res Commun 1995; 217(2): 624-31.
Dehrmann FM, Coetzer TH, Pike RN, Dennison C. Mature cathepsin L is substantially active in the ionic milieu of the extracellular medium. Arch Biochem Biophys 1995; 324(1): 93-8.
Sudhan DR, Siemann DW. Cathepsin L inhibition by the small molecule KGP94 suppresses tumor microenvironment enhanced metastasis associated cell functions of prostate and breast cancer cells. Clin Exp Metastasis 2013; 30(7): 891-902.
Guo Z, Huo J, Di J, Zeng S, Liu J, Xing F. PI3K pathway inhibitor LY294002 alters Jurkat T cell biobehaviours via ERK1/2-ICBP90 mediation. Cent Eur J Biol 2014; 9(8): 739-48.
Nishida Y, Kohno K, Kawamata T, Morimitsu K, Kuwano M, Miyakawa I. Increased cathepsin L levels in serum in some patients with ovarian cancer: comparison with CA125 and CA72-4. Gynecol Oncol 1995; 56(3): 357-61.
Chambers AF, Colella R, Denhardt DT, Wilson SM. Increased expression of cathepsins L and B and decreased activity of their inhibitors in metastatic, ras-transformed NIH 3T3 cells. Mol Carcinog 1992; 5(3): 238-45.
Frade R, Rodrigues-Lima F, Huang S, Xie K, Guillaume N, Bar-Eli M. Procathepsin-L, a proteinase that cleaves human C3 (the third component of complement), confers high tumorigenic and metastatic properties to human melanoma cells. Cancer Res 1998; 58(13): 2733-6.
Yang Z, Cox JL. Cathepsin L increases invasion and migration of B16 melanoma. Cancer Cell Int 2007; 7: 8.
Chung JH. Im, E.K.; Jin, T.W.; Lee, S.M.; Kim, S.H.; Choi, E.Y.; Shin, M.J.; Lee, K.H.; Jang, Y. Cathepsin L derived from skeletal muscle cells transfected with bFGF promotes endothelial cell migration. Exp Mol Med 2011; 43(4): 179-88.
Urbich C, Heeschen C, Aicher A, et al. Cathepsin L is required for endothelial progenitor cell-induced neovascularization. Nat Med 2005; 11(2): 206-13.
Shimada N, Ohno-Matsui K, Iseki S, et al. Cathepsin L in bone marrow-derived cells is required for retinal and choroidal neovascularization. Am J Pathol 2010; 176(5): 2571-80.
Felbor U, Dreier L, Bryant RA, Ploegh HL, Olsen BR, Mothes W. Secreted cathepsin L generates endostatin from collagen XVIII. EMBO J 2000; 19(6): 1187-94.
Gocheva V, Zeng W, Ke D, et al. Distinct roles for cysteine cathepsin genes in multistage tumorigenesis. Genes Dev 2006; 20(5): 543-56.

Rights & PermissionsPrintExport Cite as

Article Details

Year: 2019
Page: [231 - 242]
Pages: 12
DOI: 10.2174/1568009618666180831123951
Price: $58

Article Metrics

PDF: 25