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Breast Cancer Research
Published in: Breast Cancer Research 1/2013

Open Access 01-02-2013 | Research article

Anti-matrix metalloproteinase-9 DNAzyme decreases tumor growth in the MMTV-PyMT mouse model of breast cancer

Authors: Miranda A Hallett, Bin Teng, Hisashi Hasegawa, Luciana P Schwab, Tiffany N Seagroves, Tayebeh Pourmotabbed

Published in: Breast Cancer Research | Issue 1/2013

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Abstract

Introduction

Despite continued improvements in diagnosis, surgical techniques, and chemotherapy, breast cancer patients are still overcome by cancer metastasis. Tumor cell proliferation, invasion and metastasis are mediated, at least in part, through degradation of basement membrane by neutral matrix metalloproteinases (MMP) produced by tumor and stromal cells. Evidence suggests that MMP-9 plays a significant role in breast tumor cell invasion and metastasis. DNAzymes or catalytic oligonucleotides are new classes of gene targeting molecules that bind and cleave a specific mRNA, resulting in decreased protein expression.

Methods

The application of anti-MMP-9 DNAzyme (AM9D) for the treatment of primary and metastatic breast cancer was evaluated in vitro and in vivo using MDA-MB-231 cells and the MMTV-PyMT transgenic breast cancer mouse model. Spontaneously developed mammary tumors in MMTV-PyMT transgenic mice were treated intratumorally with naked AM9D, once a week for 4 weeks. The stability of DNAzyme was determined in vitro and in vivo using fluorescently labeled DNAzyme.

Results

AM9D specifically inhibited expression of MMP-9 in MDA-MB-231 cells resulting in reduced invasive property of these cells by 43%. Weekly intratumoral treatment of spontaneously developed mammary tumors in MMTV-PyMT transgenic mice was sufficient to significantly reduce the rate of tumor growth and final tumor load in a dose dependent and statistically significant manner (P < 0.05). This decrease in tumor growth was correlated with decreased MMP-9 protein production within the treated tumor tissues. Tumors treated with AM9D were also less vascularized and contained more apoptotic cells compared to control and untreated tumors.

Conclusions

These results show that targeting and down regulation of MMP-9 by AM9D could prove useful as a therapy against breast carcinoma tumor growth and invasion.
Appendix
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Literature
1.
2.
Weigelt B, Peterse JL, van 't Veer LJ: Breast cancer metastasis: markers and models. Nat Rev Cancer. 2005, 5: 591-602.CrossRefPubMed
3.
Phadke PA, Vaidya KS, Nash KT, Hurst DR, Welch DR: BRMS1 suppresses breast cancer experimental metastasis to multiple organs by inhibiting several steps of the metastatic process. Am J Pathol. 2008, 172: 809-817. 10.2353/ajpath.2008.070772.CrossRefPubMedPubMedCentral
4.
Sethi N, Kang Y: Unravelling the complexity of metastasis - molecular understanding and targeted therapies. Nat Rev Cancer. 2011, 11: 735-748. 10.1038/nrc3125.CrossRefPubMed
5.
Chambers AF, Groom AC, MacDonald IC: Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer. 2002, 2: 563-572. 10.1038/nrc865.CrossRefPubMed
6.
Gavrilovic J, Moens G, Thiery JP, Jouanneau J: Expression of transfected transforming growth factor alpha induces a motile fibroblast-like phenotype with extracellular matrix-degrading potential in a rat bladder carcinoma cell line. Cell Regul. 1990, 1: 1003-1014.PubMedPubMedCentral
7.
Lochter A, Galosy S, Muschler J, Freedman N, Werb Z, Bissell MJ: Matrix metalloproteinase stromelysin-1 triggers a cascade of molecular alterations that leads to stable epithelial-to-mesenchymal conversion and a premalignant phenotype in mammary epithelial cells. J Cell Biol. 1997, 139: 1861-1872. 10.1083/jcb.139.7.1861.CrossRefPubMedPubMedCentral
8.
Noel A, Jost M, Maquoi E: Matrix metalloproteinases at cancer tumor-host interface. Semin Cell Dev Biol. 2008, 19: 52-60. 10.1016/j.semcdb.2007.05.011.CrossRefPubMed
9.
Stuelten CH, DaCosta Byfield S, Arany PR, Karpova TS, Stetler-Stevenson WG, Roberts AB: Breast cancer cells induce stromal fibroblasts to express MMP-9 via secretion of TNF-alpha and TGF-beta. J Cell Sci. 2005, 118: 2143-2153. 10.1242/jcs.02334.CrossRefPubMed
10.
Egeblad M, Werb Z: New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer. 2002, 2: 161-174. 10.1038/nrc745.CrossRefPubMed
11.
Watanabe H, Nakanishi I, Yamashita K, Hayakawa T, Okada Y: Matrix metalloproteinase-9 (92 kDa gelatinase/type IV collagenase) from U937 monoblastoid cells: correlation with cellular invasion. J Cell Sci. 1993, 104 (Pt 4): 991-999.PubMed
12.
Cha HJ, Bae SK, Lee HY, Lee OH, Sato H, Seiki M, Park BC, Kim KW: Anti-invasive activity of ursolic acid correlates with the reduced expression of matrix metalloproteinase-9 (MMP-9) in HT1080 human fibrosarcoma cells. Cancer Res. 1996, 56: 2281-2284.PubMed
13.
Fenhalls G, Geyp M, Dent DM, Parker MI: Breast tumour cell-induced down-regulation of type I collagen mRNA in fibroblasts. Br J Cancer. 1999, 81: 1142-1149. 10.1038/sj.bjc.6690821.CrossRefPubMedPubMedCentral
14.
Coussens LM, Werb Z: Matrix metalloproteinases and the development of cancer. Chem Biol. 1996, 3: 895-904. 10.1016/S1074-5521(96)90178-7.CrossRefPubMed
15.
Rao JS: Molecular mechanisms of glioma invasiveness: the role of proteases. Nat Rev Cancer. 2003, 3: 489-501. 10.1038/nrc1121.CrossRefPubMed
16.
Hojilla CV, Mohammed FF, Khokha R: Matrix metalloproteinases and their tissue inhibitors direct cell fate during cancer development. Br J Cancer. 2003, 89: 1817-1821. 10.1038/sj.bjc.6601327.CrossRefPubMedPubMedCentral
17.
Bhoopathi P, Chetty C, Kunigal S, Vanamala SK, Rao JS, Lakka SS: Blockade of tumor growth due to matrix metalloproteinase-9 inhibition is mediated by sequential activation of beta1-integrin, ERK, and NF-kappaB. J Biol Chem. 2008, 283: 1545-1552.CrossRefPubMed
18.
Matrisian LM, Bowden GT, Krieg P, Furstenberger G, Briand JP, Leroy P, Breathnach R: The mRNA coding for the secreted protease transin is expressed more abundantly in malignant than in benign tumors. Proc Natl Acad Sci USA. 1986, 83: 9413-9417. 10.1073/pnas.83.24.9413.CrossRefPubMedPubMedCentral
19.
Liotta LA, Rao CN, Wewer UM: Biochemical interactions of tumor cells with the basement membrane. Annu Rev Biochem. 1986, 55: 1037-1057. 10.1146/annurev.bi.55.070186.005133.CrossRefPubMed
20.
Chintala SK, Tonn JC, Rao JS: Matrix metalloproteinases and their biological function in human gliomas. Int J Dev Neurosci. 1999, 17: 495-502. 10.1016/S0736-5748(99)00010-6.CrossRefPubMed
21.
Morini M, Mottolese M, Ferrari N, Ghiorzo F, Buglioni S, Mortarini R, Noonan DM, Natali PG, Albini A: The alpha 3 beta 1 integrin is associated with mammary carcinoma cell metastasis, invasion, and gelatinase B (MMP-9) activity. Int J Cancer. 2000, 87: 336-342. 10.1002/1097-0215(20000801)87:3<336::AID-IJC5>3.0.CO;2-3.CrossRefPubMed
22.
Bernhard EJ, Gruber SB, Muschel RJ: Direct evidence linking expression of matrix metalloproteinase 9 (92-kDa gelatinase/collagenase) to the metastatic phenotype in transformed rat embryo cells. Proc Natl Acad Sci USA. 1994, 91: 4293-4297. 10.1073/pnas.91.10.4293.CrossRefPubMedPubMedCentral
23.
Santoro SW, Joyce GF: Mechanism and utility of an RNA-cleaving DNA enzyme. Biochemistry. 1998, 37: 13330-13342. 10.1021/bi9812221.CrossRefPubMed
24.
25.
Dass CR, Choong PF, Khachigian LM: DNAzyme technology and cancer therapy: cleave and let die. Mol Cancer Ther. 2008, 7: 243-251.CrossRefPubMed
26.
Lowe HC, Fahmy RG, Kavurma MM, Baker A, Chesterman CN, Khachigian LM: Catalytic oligodeoxynucleotides define a key regulatory role for early growth response factor-1 in the porcine model of coronary in-stent restenosis. Circ Res. 2001, 89: 670-677. 10.1161/hh2001.097867.CrossRefPubMed
27.
Khachigian LM, Fahmy RG, Zhang G, Bobryshev YV, Kaniaros A: c-Jun regulates vascular smooth muscle cell growth and neointima formation after arterial injury. Inhibition by a novel DNA enzyme targeting c-Jun. J Biol Chem. 2002, 277: 22985-22991. 10.1074/jbc.M200977200.CrossRefPubMed
28.
Zhang L, Gasper WJ, Stass SA, Ioffe OB, Davis MA, Mixson AJ: Angiogenic inhibition mediated by a DNAzyme that targets vascular endothelial growth factor receptor 2. Cancer Res. 2002, 62: 5463-5469.PubMed
29.
Sun LQ, Cairns MJ, Saravolac EG, Baker A, Gerlach WL: Catalytic nucleic acids: from lab to applications. Pharmacol Rev. 2000, 52: 325-347.PubMed
30.
Hegedüs L, Cho H, Xie X, Eliceiri GL: Additional MDA-MB-231 breast cancer cell matrix metalloproteinases promote invasiveness. J Cell Physiol. 2008, 216: 480-485. 10.1002/jcp.21417.CrossRefPubMed
31.
Bu CH, Pourmotabbed T: Mechanism of activation of human neutrophil gelatinase B: Discriminating between the role of Ca in activation and catalytsis. J Biol Chem. 1995, 18563-18569. 270
32.
Chantrain CF, DeClerck YA, Groshen S, McNamara G: Computerized quantification of tissue vascularization using high-resolution slide scanning of whole tumor sections. J Histochem Cytochem. 2003, 51: 151-158. 10.1177/002215540305100203.CrossRefPubMed
33.
Yodkeeree S, Ampasavate C, Sung B, Aggarwal BB, Limtrakul P: Demethoxycurcumin suppresses migration and invasion of MDA-MB-231 human breast cancer cell line. Eur J Pharmacol. 2010, 627: 8-15. 10.1016/j.ejphar.2009.09.052.CrossRefPubMed
34.
Lin EY, Jones JG, Li P, Zhu L, Whitney KD, Muller WJ, Pollard JW: Progression to malignancy in the polyoma middle T oncoprotein mouse breast cancer model provides a reliable model for human diseases. Am J Pathol. 2003, 163: 2113-2126. 10.1016/S0002-9440(10)63568-7.CrossRefPubMedPubMedCentral
35.
36.
Herschkowitz JI, Simin K, Weigman VJ, Mikaelian I, Usary J, Hu Z, Rasmussen KE, Jones LP, Assefnia S, Chandrasekharan S, Backlund MG, Yin Y, Khramtsov IA, Bastein R, Quackenbush J, Glazer RI, Brown PH, Green JE, Kopelovich L, Furth PA, Palazzo JP, Olopade OI, Bernard PS, Churchill GA, Dyke TV, Perou CM: Identification of conserved gene expression features between murine mammary carcinoma models and human breast tumors. Genome Biol. 2007, 8: R76-10.1186/gb-2007-8-5-r76.CrossRefPubMedPubMedCentral
37.
Pedersen TX, Pennington CJ, Almholt K, Christensen IJ, Nielsen BS, Edwards DR, Romer J, Dano K, Johnsen M: Extracellular protease mRNAs are predominantly expressed in the stromal areas of microdissected mouse breast carcinomas. Carcinogenesis. 2005, 26: 1233-1240.CrossRefPubMed
38.
Szabova L, Yamada SS, Birkedal-Hansen H, Holmbeck K: Expression pattern of four membrane-type matrix metalloproteinases in the normal and diseased mouse mammary gland. J Cell Physiol. 2005, 205: 123-132. 10.1002/jcp.20385.CrossRefPubMed
39.
Almholt K, Green KA, Juncker-Jensen A, Nielsen BS, Lund LR, Romer J: Extracellular proteolysis in transgenic mouse models of breast cancer. J Mammary Gland Biol Neoplasia. 2007, 12: 83-97. 10.1007/s10911-007-9040-x.CrossRefPubMedPubMedCentral
40.
Almholt K, Juncker-Jensen A, Laerum OD, Dano K, Johnsen M, Lund LR, Romer J: Metastasis is strongly reduced by the matrix metalloproteinase inhibitor Galardin in the MMTV-PymT transgenic breast cancer model. Mol Cancer Ther. 2008, 7: 2758-2767. 10.1158/1535-7163.MCT-08-0251.CrossRefPubMed
41.
Lin EY, Nguyen AV, Russell RG, Pollard JW: Colony-stimulating factor 1 promotes progression of mammary tumors to malignancy. J Exp Med. 2001, 193: 727-740. 10.1084/jem.193.6.727.CrossRefPubMedPubMedCentral
42.
Lin EY, Gouon-Evans V, Nguyen AV, Pollard JW: The macrophage growth factor CSF-1 in mammary gland development and tumor progression. J Mammary Gland Biol Neoplasia. 2002, 7: 147-162. 10.1023/A:1020399802795.CrossRefPubMed
43.
Bergers G, Brekken R, McMahon G, Vu TH, Itoh T, Tamaki K, Tanzawa K, Thorpe P, Itohara S, Werb Z, Hanahan D: Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. Nat Cell Biol. 2000, 2: 737-744. 10.1038/35036374.CrossRefPubMedPubMedCentral
44.
Martin MD, Carter KJ, Jean-Philippe SR, Chang M, Mobashery S, Thiolloy S, Lynch CC, Matrisian LM, Fingleton B: Effect of ablation or inhibition of stromal matrix metalloproteinase-9 on lung metastasis in a breast cancer model is dependent on genetic background. Cancer Res. 2008, 68: 6251-6259. 10.1158/0008-5472.CAN-08-0537.CrossRefPubMedPubMedCentral
45.
46.
Wojtowicz-Praga SM, Dickson RB, Hawkins MJ: Matrix metalloproteinase inhibitors. Invest New Drugs. 1997, 15: 61-75. 10.1023/A:1005722729132.CrossRefPubMed
47.
Rasmussen HS, McCann PP: Matrix metalloproteinase inhibition as a novel anticancer strategy: a review with special focus on batimastat and marimastat. Pharmacol Ther. 1997, 75: 69-75. 10.1016/S0163-7258(97)00023-5.CrossRefPubMed
48.
Martin MD, Matrisian LM: The other side of MMPs: protective roles in tumor progression. Cancer Metastasis Rev. 2007, 26: 717-724. 10.1007/s10555-007-9089-4.CrossRefPubMed
49.
Coussens LM, Fingleton B, Matrisian LM: Matrix metalloproteinase inhibitors and cancer: trials and tribulations. Science. 2002, 295: 2387-2392. 10.1126/science.1067100.CrossRefPubMed
Metadata
Title
Anti-matrix metalloproteinase-9 DNAzyme decreases tumor growth in the MMTV-PyMT mouse model of breast cancer
Authors
Miranda A Hallett
Bin Teng
Hisashi Hasegawa
Luciana P Schwab
Tiffany N Seagroves
Tayebeh Pourmotabbed
Publication date
01-02-2013
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 1/2013
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr3385

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