Suppression of Colorectal Cancer Liver Metastasis by Apolipoprotein(a) Kringle V in a Nude Mouse Model through the Induction of Apoptosis in Tumor-Associated Endothelial Cells
The formation of liver metastases in colorectal cancer patients is the primary cause of patient death. Current therapies directed at liver metastasis from colorectal cancer have had minimal impact on patient outcomes. Therefore, the development of alternative treatment strategies for liver metastasis is needed. In the present study, we demonstrated that recombinant human apolipoprotein(a) kringle V, also known as rhLK8, induced the apoptotic turnover of endothelial cells in vitro through the mitochondrial apoptosis pathway. The interaction of rhLK8 with glucose-regulated protein 78 (GRP78) may be involved in the induction of apoptosis because the inhibition of GRP78 by GRP78-specific antibodies or siRNA knockdown inhibited the rhLK8-mediated apoptosis of human umbilical vein endothelial cells in vitro. Next, to evaluate the effects of rhLK8 on angiogenesis and metastasis, an experimental model of liver metastasis was established by injecting a human colorectal cancer cell line, LS174T, into the spleens of BALB/c nude mice. The systemic administration of rhLK8 significantly suppressed liver metastasis from human colorectal cancer cells and improved host survival compared with controls. The combination of rhLK8 and 5-fluorouracil substantially increased these survival benefits compared with either therapy alone. Histological observation showed significant induction of apoptosis among tumor-associated endothelial cells in liver metastases from rhLK8-treated mice compared with control mice. Collectively, these results suggest that the combination of rhLK8 with conventional chemotherapy may be a promising approach for the treatment of patients with life-threatening colorectal cancer liver metastases.
References
[1]
Siegel R, Naishadham D, Jemal A (2013) Cancer statistics, 2013. CA Cancer J Clin 63: 11–30. doi: 10.3322/caac.21166
Scheele J, Stangl R, Altendorf-Hofmann A (1990) Hepatic metastases from colorectal carcinoma: impact of surgical resection on the natural history. Br J Surg 77: 1241–1246. doi: 10.1002/bjs.1800771115
[4]
Lorenz M, Staib-Sebler E, Hochmuth K, Heinrich S, Gog C, et al. (2000) Surgical resection of liver metastases of colorectal carcinoma: short and long-term results. Semin Oncol 27: 112–119.
[5]
Folkman J (1971) Tumor angiogenesis: therapeutic implications. N Engl J Med 285: 1182–1186. doi: 10.1056/nejm197111182852108
[6]
Folkman J (1990) What is the evidence that tumors are angiogenesis dependent? J Natl Cancer Inst 82: 4–6. doi: 10.1093/jnci/82.1.4
[7]
O’Reilly MS, Holmgren L, Chen C, Folkman J (1996) Angiostatin induces and sustains dormancy of human primary tumors in mice. Nat Med 2: 689–692. doi: 10.1038/nm0696-689
[8]
te Velde EA, Reijerkerk A, Brandsma D, Vogten JM, Wu Y, et al. (2005) Early endostatin treatment inhibits metastatic seeding of murine colorectal cancer cells in the liver and their adhesion to endothelial cells. Br J Cancer 92: 729–735.
[9]
Takeda A, Stoeltzing O, Ahmad SA, Reinmuth N, Liu W, et al. (2002) Role of angiogenesis in the development and growth of liver metastasis. Ann Surg Oncol 9: 610–616. doi: 10.1007/bf02574475
[10]
Chambers AF, Groom AC, MacDonald IC (2002) Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer 2: 563–572. doi: 10.1038/nrc865
[11]
Chambers AF, MacDonald IC, Schmidt EE, Morris VL, Groom AC (2000) Clinical targets for anti-metastasis therapy. Adv Cancer Res 79: 91–121. doi: 10.1016/s0065-230x(00)79003-8
[12]
Katsouras CS, Karabina SA, Tambaki AP, Goudevenos JA, Michalis LK, et al. (2001) Serum lipoprotein(a) concentrations and apolipoprotein(a) isoforms: association with the severity of clinical presentation in patients with coronary heart disease. J Cardiovasc Risk 8: 311–317. doi: 10.1177/174182670100800511
[13]
McLean JW, Tomlinson JE, Kuang WJ, Eaton DL, Chen EY, et al. (1987) cDNA sequence of human apolipoprotein(a) is homologous to plasminogen. Nature 330: 132–137. doi: 10.1038/330132a0
[14]
Cao Y, Cao R, Veitonmaki N (2002) Kringle structures and antiangiogenesis. Curr Med Chem Anticancer Agents 2: 667–681. doi: 10.2174/1568011023353705
[15]
Schulter V, Koolwijk P, Peters E, Frank S, Hrzenjak A, et al. (2001) Impact of apolipoprotein(a) on in vitro angiogenesis. Arterioscler Thromb Vasc Biol 21: 433–438. doi: 10.1161/01.atv.21.3.433
[16]
Trieu VN, Uckun FM (1999) Apolipoprotein(a), a link between atherosclerosis and tumor angiogenesis. Biochem Biophys Res Commun 257: 714–718. doi: 10.1006/bbrc.1999.0519
[17]
Kim JS, Chang JH, Yu HK, Ahn JH, Yum JS, et al. (2003) Inhibition of angiogenesis and angiogenesis-dependent tumor growth by the cryptic kringle fragments of human apolipoprotein(a). J Biol Chem 278: 29000–29008. doi: 10.1074/jbc.m301042200
[18]
Kim JS, Yu HK, Ahn JH, Lee HJ, Hong SW, et al. (2004) Human apolipoprotein(a) kringle V inhibits angiogenesis in vitro and in vivo by interfering with the activation of focal adhesion kinases. Biochem Biophys Res Commun 313: 534–540. doi: 10.1016/j.bbrc.2003.11.148
[19]
Lee HJ, Yu HK, Papadopoulos JN, Kim SW, He J, et al. (2012) Targeted antivascular therapy with the apolipoprotein(a) kringle V, rhLK8, inhibits the growth and metastasis of human prostate cancer in an orthotopic nude mouse model. Neoplasia 14: 335–343.
[20]
Lee TH, Kim MD, Shin SY, Lim HK, Seo JH (2006) Disruption of hexokinase II (HXK2) partly relieves glucose repression to enhance production of human kringle fragment in gratuitous recombinant Saccharomyces cerevisiae. J Biotechnol 126: 562–567. doi: 10.1016/j.jbiotec.2006.05.011
[21]
Kang KY, Park JH, Lim IH, Kim SG, Park SH, et al. (2006) Crystallization of antiangiogenic Kringle V derived from human apolipoprotein A: crystallization applied to purification and formulation. Biosci Biotechnol Biochem 70: 916–925. doi: 10.1271/bbb.70.916
[22]
Pijpers AH, van Setten PA, van den Heuvel LP, Assmann KJ, Dijkman HB, et al. (2001) Verocytotoxin-induced apoptosis of human microvascular endothelial cells. J Am Soc Nephrol 12: 767–778.
[23]
Mikhailov V, Mikhailova M, Pulkrabek DJ, Dong Z, Venkatachalam MA, et al. (2001) Bcl-2 prevents Bax oligomerization in the mitochondrial outer membrane. J Biol Chem 276: 18361–18374. doi: 10.1074/jbc.m100655200
[24]
Kim SJ, Uehara H, Yazici S, Langley RR, He J, et al. (2004) Simultaneous blockade of platelet-derived growth factor-receptor and epidermal growth factor-receptor signaling and systemic administration of paclitaxel as therapy for human prostate cancer metastasis in bone of nude mice. Cancer Res 64: 4201–4208. doi: 10.1158/0008-5472.can-03-3763
[25]
Mow BM, Blajeski AL, Chandra J, Kaufmann SH (2001) Apoptosis and the response to anticancer therapy. Curr Opin Oncol 13: 453–462. doi: 10.1097/00001622-200111000-00007
[26]
Davidson DJ, Haskell C, Majest S, Kherzai A, Egan DA, et al. (2005) Kringle 5 of human plasminogen induces apoptosis of endothelial and tumor cells through surface-expressed glucose-regulated protein 78. Cancer Res 65: 4663–4672. doi: 10.1158/0008-5472.can-04-3426
[27]
Fidler IJ, Hart IR (1982) Biological diversity in metastatic neoplasms: origins and implications. Science 217: 998–1003. doi: 10.1126/science.7112116
[28]
Hofmeister V, Schrama D, Becker JC (2008) Anti-cancer therapies targeting the tumor stroma. Cancer Immunol Immunother 57: 1–17. doi: 10.1007/s00262-007-0365-5
[29]
Jung YD, Ahmad SA, Akagi Y, Takahashi Y, Liu W, et al. (2000) Role of the tumor microenvironment in mediating response to anti-angiogenic therapy. Cancer Metastasis Rev 19: 147–157.
[30]
Ferrara N, Hillan KJ, Gerber HP, Novotny W (2004) Discovery and development of bevacizumab, an anti-VEGF antibody for treating cancer. Nat Rev Drug Discov 3: 391–400. doi: 10.1038/nrd1381
[31]
Hobson B, Denekamp J (1984) Endothelial proliferation in tumours and normal tissues: continuous labelling studies. Br J Cancer 49: 405–413.
[32]
Eberhard A, Kahlert S, Goede V, Hemmerlein B, Plate KH, et al. (2000) Heterogeneity of angiogenesis and blood vessel maturation in human tumors: implications for antiangiogenic tumor therapies. Cancer Res 60: 1388–1393.
[33]
Chavakis E, Dimmeler S (2002) Regulation of endothelial cell survival and apoptosis during angiogenesis. Arterioscler Thromb Vasc Biol 22: 887–893. doi: 10.1161/01.atv.0000017728.55907.a9
[34]
Hanahan D, Folkman J (1996) Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell 86: 353–364. doi: 10.1016/s0092-8674(00)80108-7
[35]
Lucas R, Holmgren L, Garcia I, Jimenez B, Mandriota SJ, et al. (1998) Multiple forms of angiostatin induce apoptosis in endothelial cells. Blood 92: 4730–4741.
[36]
Cao Y, Chen A, An SS, Ji RW, Davidson D, et al. (1997) Kringle 5 of plasminogen is a novel inhibitor of endothelial cell growth. J Biol Chem 272: 22924–22928. doi: 10.1074/jbc.272.36.22924
[37]
Ji WR, Castellino FJ, Chang Y, Deford ME, Gray H, et al. (1998) Characterization of kringle domains of angiostatin as antagonists of endothelial cell migration, an important process in angiogenesis. FASEB J 12: 1731–1738.
[38]
Perri SR, Nalbantoglu J, Annabi B, Koty Z, Lejeune L, et al. (2005) Plasminogen kringle 5-engineered glioma cells block migration of tumor-associated macrophages and suppress tumor vascularization and progression. Cancer Res 65: 8359–8365. doi: 10.1158/0008-5472.can-05-0508
[39]
Yang X, Cheng R, Li C, Cai W, Ma JX, et al. (2006) Kringle 5 of human plasminogen suppresses hepatocellular carcinoma growth both in grafted and xenografted mice by anti-angiogenic activity. Cancer Biol Ther 5: 399–405. doi: 10.4161/cbt.5.4.2511
[40]
Perri SR, Martineau D, Francois M, Lejeune L, Bisson L, et al. (2007) Plasminogen Kringle 5 blocks tumor progression by antiangiogenic and proinflammatory pathways. Mol Cancer Ther 6: 441–449. doi: 10.1158/1535-7163.mct-06-0434
[41]
Jin GH, Ma DY, Wu N, Marikar FM, Jin SZ, et al. (2007) Combination of human plasminogen kringle 5 with ionizing radiation significantly enhances the efficacy of antitumor effect. Int J Cancer 121: 2539–2546. doi: 10.1002/ijc.22708
[42]
Fan JK, Xiao T, Gu JF, Wei N, He LF, et al. (2008) Increased suppression of oncolytic adenovirus carrying mutant k5 on colorectal tumor. Biochem Biophys Res Commun 374: 198–203. doi: 10.1016/j.bbrc.2008.07.005
[43]
Li Y, Han W, Zhang Y, Yuan L, Shi X, et al. (2008) Intramuscular electroporation of a plasmid encoding human plasminogen kringle 5 induces growth inhibition of Lewis lung carcinoma in mice. Cancer Biother Radiopharm 23: 332–341. doi: 10.1089/cbr.2007.0437
[44]
Nguyen TM, Subramanian IV, Kelekar A, Ramakrishnan S (2007) Kringle 5 of human plasminogen, an angiogenesis inhibitor, induces both autophagy and apoptotic death in endothelial cells. Blood 109: 4793–4802. doi: 10.1182/blood-2006-11-059352
[45]
Gu X, Yao Y, Cheng R, Zhang Y, Dai Z, et al. (2011) Plasminogen K5 activates mitochondrial apoptosis pathway in endothelial cells by regulating Bak and Bcl-x(L) subcellular distribution. Apoptosis 16: 846–855. doi: 10.1007/s10495-011-0618-9
[46]
Cory S, Adams JM (2002) The Bcl2 family: regulators of the cellular life-or-death switch. Nat Rev Cancer 2: 647–656. doi: 10.1038/nrc883
[47]
Gerber HP, Ferrara N (2005) Pharmacology and pharmacodynamics of bevacizumab as monotherapy or in combination with cytotoxic therapy in preclinical studies. Cancer Res 65: 671–680.
[48]
Jain RK (2005) Normalization of tumor vasculature: an emerging concept in antiangiogenic therapy. Science 307: 58–62. doi: 10.1126/science.1104819
[49]
Ma J, Waxman DJ (2008) Combination of antiangiogenesis with chemotherapy for more effective cancer treatment. Mol Cancer Ther 7: 3670–3684. doi: 10.1158/1535-7163.mct-08-0715
[50]
Huang G, Chen L (2008) Tumor vasculature and microenvironment normalization: a possible mechanism of antiangiogenesis therapy. Cancer Biother Radiopharm 23: 661–667. doi: 10.1089/cbr.2008.0492
