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BMC Cancer
Published in: BMC Cancer 1/2014

Open Access 01-12-2014 | Research article

Identification of genes regulating migration and invasion using a new model of metastatic prostate cancer

Authors: Jacqueline Banyard, Ivy Chung, Matthew Migliozzi, Derek T Phan, Arianne M Wilson, Bruce R Zetter, Diane R Bielenberg

Published in: BMC Cancer | Issue 1/2014

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Abstract

Background

Understanding the complex, multistep process of metastasis remains a major challenge in cancer research. Metastasis models can reveal insights in tumor development and progression and provide tools to test new intervention strategies.

Methods

To develop a new cancer metastasis model, we used DU145 human prostate cancer cells and performed repeated rounds of orthotopic prostate injection and selection of subsequent lymph node metastases. Tumor growth, metastasis, cell migration and invasion were analyzed. Microarray analysis was used to identify cell migration- and cancer-related genes correlating with metastasis. Selected genes were silenced using siRNA, and their roles in cell migration and invasion were determined in transwell migration and Matrigel invasion assays.

Results

Our in vivo cycling strategy created cell lines with dramatically increased tumorigenesis and increased ability to colonize lymph nodes (DU145LN1-LN4). Prostate tumor xenografts displayed increased vascularization, enlarged podoplanin-positive lymphatic vessels and invasive margins. Microarray analysis revealed gene expression profiles that correlated with metastatic potential. Using gene network analysis we selected 3 significantly upregulated cell movement and cancer related genes for further analysis: EPCAM (epithelial cell adhesion molecule), ITGB4 (integrin β4) and PLAU (urokinase-type plasminogen activator (uPA)). These genes all showed increased protein expression in the more metastatic DU145-LN4 cells compared to the parental DU145. SiRNA knockdown of EpCAM, integrin-β4 or uPA all significantly reduced cell migration in DU145-LN4 cells. In contrast, only uPA siRNA inhibited cell invasion into Matrigel. This role of uPA in cell invasion was confirmed using the uPA inhibitors, amiloride and UK122.

Conclusions

Our approach has identified genes required for the migration and invasion of metastatic tumor cells, and we propose that our new in vivo model system will be a powerful tool to interrogate the metastatic cascade in prostate cancer.
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Literature
1.
American Cancer Society: Cancer Facts & Figures 2012. 2012, Atlanta: American Cancer Society
2.
Siegel R, DeSantis C, Virgo K, Stein K, Mariotto A, Smith T, Cooper D, Gansler T, Lerro C, Fedewa S, Lin C, Leach C, Cannady RS, Cho H, Scoppa S, Hachey M, Kirch R, Jemal A, Ward E: Cancer treatment and survivorship statistics, 2012. CA Cancer J Clin. 2012, 62 (4): 220-241. 10.3322/caac.21149.CrossRefPubMed
3.
Smith JA, Seaman JP, Gleidman JB, Middleton RG: Pelvic lymph node metastasis from prostatic cancer: influence of tumor grade and stage in 452 consecutive patients. J Urol. 1983, 130 (2): 290-292.PubMed
4.
Dadras SS, Paul T, Bertoncini J, Brown LF, Muzikansky A, Jackson DG, Ellwanger U, Garbe C, Mihm MC, Detmar M: Tumor lymphangiogenesis: a novel prognostic indicator for cutaneous melanoma metastasis and survival. Am J Pathol. 2003, 162 (6): 1951-1960. 10.1016/S0002-9440(10)64328-3.CrossRefPubMedPubMedCentral
5.
Cheng L, Jones TD, Lin H, Eble JN, Zeng G, Carr MD, Koch MO: Lymphovascular invasion is an independent prognostic factor in prostatic adenocarcinoma. J Urol. 2005, 174 (6): 2181-2185. 10.1097/01.ju.0000181215.41607.c3.CrossRefPubMed
6.
Rinderknecht M, Detmar M: Tumor lymphangiogenesis and melanoma metastasis. J Cell Physiol. 2008, 216 (2): 347-354. 10.1002/jcp.21494.CrossRefPubMed
7.
Karakiewicz PI, Hutterer GC: Predictive models and prostate cancer. Nat Clin Pract Urol. 2008, 5 (2): 82-92. 10.1038/ncpuro0972.CrossRefPubMed
8.
Sleeman JP, Thiele W: Tumor metastasis and the lymphatic vasculature. Int J Cancer. 2009, 125 (12): 2747-2756. 10.1002/ijc.24702.CrossRefPubMed
9.
10.
Wilt TJ, Brawer MK, Jones KM, Barry MJ, Aronson WJ, Fox S, Gingrich JR, Wei JT, Gilhooly P, Grob BM, Nsouli I, Iyer P, Cartagena R, Snider G, Roehrborn C, Sharifi R, Blank W, Pandya P, Andriole GL, Culkin D, Wheeler T, Prostate Cancer Intervention versus Observation Trial (PIVOT) Study Group: Radical prostatectomy versus observation for localized prostate cancer. N Engl J Med. 2012, 367 (3): 203-213. 10.1056/NEJMoa1113162.CrossRefPubMedPubMedCentral
11.
Aizer AA, Paly JJ, Zietman AL, Nguyen PL, Beard CJ, Rao SK, Kaplan ID, Niemierko A, Hirsch MS, Wu CL, Olumi AF, Michaelson MD, D'Amico AV, Efstathiou JA: Multidisciplinary care and pursuit of active surveillance in low-risk prostate cancer. J Clin Oncol. 2012, 30 (25): 3071-3076. 10.1200/JCO.2012.42.8466.CrossRefPubMed
12.
Stephenson RA, Dinney CP, Gohji K, Ordonez NG, Killion JJ, Fidler IJ: Metastatic model for human prostate cancer using orthotopic implantation in nude mice. J Natl Cancer Inst. 1992, 84 (12): 951-957. 10.1093/jnci/84.12.951.CrossRefPubMed
13.
Perrotte PJR, Bielenberg DR, Eve BY, Dinney CPN: Organ-specific angiogenesis and metastasis of human bladder carcinoma growing in athymic mice. Mol Urol. 1997, 1 (4): 299-307.
14.
Bielenberg DR, Fidler IJ: Regulation of Angiogenesis by the Organ Microenvironment. Antiangiogenic Agents in Cancer Therapy. Edited by: Teicher BA. 1999, Totowa: Humana Press, 77-91. 6CrossRef
15.
Fidler IJ: The organ microenvironment and cancer metastasis. Differentiation. 2002, 70 (9–10): 498-505.CrossRefPubMed
16.
Hanahan D, Weinberg RA: Hallmarks of cancer: the next generation. Cell. 2011, 144 (5): 646-674. 10.1016/j.cell.2011.02.013.CrossRefPubMed
17.
18.
Pettaway CA, Pathak S, Greene G, Ramirez E, Wilson MR, Killion JJ, Fidler IJ: Selection of highly metastatic variants of different human prostatic carcinomas using orthotopic implantation in nude mice. Clin Cancer Res. 1996, 2 (9): 1627-1636.PubMed
19.
Sobel RE, Sadar MD: Cell lines used in prostate cancer research: a compendium of old and new lines–part 1. J Urol. 2005, 173 (2): 342-359. 10.1097/01.ju.0000141580.30910.57.CrossRefPubMed
20.
Stone KR, Mickey DD, Wunderli H, Mickey GH, Paulson DF: Isolation of a human prostate carcinoma cell line (DU 145). Int J Cancer. 1978, 21 (3): 274-281. 10.1002/ijc.2910210305.CrossRefPubMed
21.
Banyard J, Chung I, Wilson AM, Vetter G, Le Béchec A, Bielenberg DR, Zetter BR: Regulation of epithelial plasticity by miR-424 and miR-200 in a new prostate cancer metastasis model. Sci Rep. 2013, 3: 3151-CrossRefPubMedPubMedCentral
22.
Chunthapong J, Seftor EA, Khalkhali-Ellis Z, Seftor RE, Amir S, Lubaroff DM, Heidger PM, Hendrix MJ: Dual roles of E-cadherin in prostate cancer invasion. J Cell Biochem. 2004, 91 (4): 649-661. 10.1002/jcb.20032.CrossRefPubMed
23.
Yilmaz M, Christofori G: Mechanisms of motility in metastasizing cells. Mol Cancer Res. 2010, 8 (5): 629-642. 10.1158/1541-7786.MCR-10-0139.CrossRefPubMed
24.
Wong SY, Hynes RO: Lymphatic or hematogenous dissemination: how does a metastatic tumor cell decide?. Cell Cycle. 2006, 5 (8): 812-817. 10.4161/cc.5.8.2646.CrossRefPubMedPubMedCentral
25.
26.
27.
Yee DS, Shariat SF, Lowrance WT, Maschino AC, Savage CJ, Cronin AM, Scardino PT, Eastham JA: Prognostic significance of lymphovascular invasion in radical prostatectomy specimens. BJU Int. 2010, 108 (4): 502-507.CrossRefPubMedPubMedCentral
28.
Weidner N, Carroll PR, Flax J, Blumenfeld W, Folkman J: Tumor angiogenesis correlates with metastasis in invasive prostate carcinoma. Am J Pathol. 1993, 143 (2): 401-409.PubMedPubMedCentral
29.
30.
Diamond E, Lee GY, Akhtar NH, Kirby BJ, Giannakakou P, Tagawa ST, Nanus DM: Isolation and characterization of circulating tumor cells in prostate cancer. Front Oncol. 2012, 2: 131-CrossRefPubMedPubMedCentral
31.
Ni J, Cozzi PJ, Duan W, Shigdar S, Graham PH, John KH, Li Y: Role of the EpCAM (CD326) in prostate cancer metastasis and progression. Cancer Metastasis Rev. 2012, 31 (3–4): 779-791.CrossRefPubMed
32.
Went P, Vasei M, Bubendorf L, Terracciano L, Tornillo L, Riede U, Kononen J, Simon R, Sauter G, Baeuerle PA: Frequent high-level expression of the immunotherapeutic target Ep-CAM in colon, stomach, prostate and lung cancers. Br J Cancer. 2006, 94 (1): 128-135. 10.1038/sj.bjc.6602924.CrossRefPubMedPubMedCentral
33.
Poczatek RB, Myers RB, Manne U, Oelschlager DK, Weiss HL, Bostwick DG, Grizzle WE: Ep-Cam levels in prostatic adenocarcinoma and prostatic intraepithelial neoplasia. J Urol. 1999, 162 (4): 1462-1466. 10.1016/S0022-5347(05)68341-3.CrossRefPubMed
34.
Zellweger T, Ninck C, Bloch M, Mirlacher M, Koivisto PA, Helin HJ, Mihatsch MJ, Gasser TC, Bubendorf L: Expression patterns of potential therapeutic targets in prostate cancer. Int J Cancer. 2005, 113 (4): 619-628. 10.1002/ijc.20615.CrossRefPubMed
35.
Benko G, Spajic B, Kruslin B, Tomas D: Impact of the EpCAM expression on biochemical recurrence-free survival in clinically localized prostate cancer. Urol Oncol. 2013, 31 (4): 468-474. 10.1016/j.urolonc.2011.03.007.CrossRefPubMed
36.
Denzel S, Maetzel D, Mack B, Eggert C, Barr G, Gires O: Initial activation of EpCAM cleavage via cell-to-cell contact. BMC Cancer. 2009, 9: 402-10.1186/1471-2407-9-402.CrossRefPubMedPubMedCentral
37.
Thuma F, Zoller M: EpCAM-associated claudin-7 supports lymphatic spread and drug resistance in rat pancreatic cancer. Int J Cancer. 2013, 133 (4): 855-866. 10.1002/ijc.28085.CrossRefPubMed
38.
Nubel T, Preobraschenski J, Tuncay H, Weiss T, Kuhn S, Ladwein M, Langbein L, Zoller M: Claudin-7 regulates EpCAM-mediated functions in tumor progression. Mol Cancer Res. 2009, 7 (3): 285-299. 10.1158/1541-7786.MCR-08-0200.CrossRefPubMed
39.
Mercurio AM, Rabinovitz I, Shaw LM: The alpha 6 beta 4 integrin and epithelial cell migration. Curr Opin Cell Biol. 2001, 13 (5): 541-545. 10.1016/S0955-0674(00)00249-0.CrossRefPubMed
40.
Giancotti FG: Targeting integrin beta4 for cancer and anti-angiogenic therapy. Trends Pharmacol Sci. 2007, 28 (10): 506-511. 10.1016/j.tips.2007.08.004.CrossRefPubMed
41.
Yoshioka T, Otero J, Chen Y, Kim YM, Koutcher JA, Satagopan J, Reuter V, Carver B, de Stanchina E, Enomoto K, Greenberg NM, Scardino PT, Scher HI, Sawyers CL, Giancotti FG: Beta4 Integrin signaling induces expansion of prostate tumor progenitors. J Clin Invest. 2013, 123 (2): 682-699.PubMedPubMedCentral
42.
Gerson KD, Maddula VS, Seligmann BE, Shearstone JR, Khan A, Mercurio AM: Effects of beta4 integrin expression on microRNA patterns in breast cancer. Biol Open. 2012, 1 (7): 658-666. 10.1242/bio.20121628.CrossRefPubMedPubMedCentral
43.
Andreasen PA, Kjoller L, Christensen L, Duffy MJ: The urokinase-type plasminogen activator system in cancer metastasis: a review. Int J Cancer. 1997, 72 (1): 1-22. 10.1002/(SICI)1097-0215(19970703)72:1<1::AID-IJC1>3.0.CO;2-Z.CrossRefPubMed
44.
Hienert G, Kirchheimer JC, Pfluger H, Binder BR: Urokinase-type plasminogen activator as a marker for the formation of distant metastases in prostatic carcinomas. J Urol. 1988, 140 (6): 1466-1469.PubMed
45.
Shariat SF, Roehrborn CG, McConnell JD, Park S, Alam N, Wheeler TM, Slawin KM: Association of the circulating levels of the urokinase system of plasminogen activation with the presence of prostate cancer and invasion, progression, and metastasis. J Clin Oncol. 2007, 25 (4): 349-355. 10.1200/JCO.2006.05.6853.CrossRefPubMed
46.
Miyake H, Hara I, Yamanaka K, Gohji K, Arakawa S, Kamidono S: Elevation of serum levels of urokinase-type plasminogen activator and its receptor is associated with disease progression and prognosis in patients with prostate cancer. Prostate. 1999, 39 (2): 123-129. 10.1002/(SICI)1097-0045(19990501)39:2<123::AID-PROS7>3.0.CO;2-2.CrossRefPubMed
47.
Pulukuri SM, Gondi CS, Lakka SS, Jutla A, Estes N, Gujrati M, Rao JS: RNA interference-directed knockdown of urokinase plasminogen activator and urokinase plasminogen activator receptor inhibits prostate cancer cell invasion, survival, and tumorigenicity in vivo. J Biol Chem. 2005, 280 (43): 36529-36540. 10.1074/jbc.M503111200.CrossRefPubMed
48.
Conn EM, Botkjaer KA, Kupriyanova TA, Andreasen PA, Deryugina EI, Quigley JP: Comparative analysis of metastasis variants derived from human prostate carcinoma cells: roles in intravasation of VEGF-mediated angiogenesis and uPA-mediated invasion. Am J Pathol. 2009, 175 (4): 1638-1652. 10.2353/ajpath.2009.090384.CrossRefPubMedPubMedCentral
49.
Mamoune A, Kassis J, Kharait S, Kloeker S, Manos E, Jones DA, Wells A: DU145 human prostate carcinoma invasiveness is modulated by urokinase receptor (uPAR) downstream of epidermal growth factor receptor (EGFR) signaling. Exp Cell Res. 2004, 299 (1): 91-100. 10.1016/j.yexcr.2004.05.008.CrossRefPubMed
50.
O’Halloran TV, Ahn R, Hankins P, Swindell E, Mazar AP: The many spaces of uPAR: delivery of theranostic agents and nanobins to multiple tumor compartments through a single target. Theranostics. 2013, 3 (7): 496-506. 10.7150/thno.4953.CrossRefPubMedPubMedCentral
51.
Sanchez-Tillo E, de Barrios O, Siles L, Amendola PG, Darling DS, Cuatrecasas M, Castells A, Postigo A: ZEB1 Promotes invasiveness of colorectal carcinoma cells through the opposing regulation of uPA and PAI-1. Clin Cancer Res. 2013, 19 (5): 1071-1082. 10.1158/1078-0432.CCR-12-2675.CrossRefPubMed
52.
Drake JM, Barnes JM, Madsen JM, Domann FE, Stipp CS, Henry MD: ZEB1 coordinately regulates laminin-332 and {beta}4 integrin expression altering the invasive phenotype of prostate cancer cells. J Biol Chem. 2010, 285 (44): 33940-33948. 10.1074/jbc.M110.136044.CrossRefPubMedPubMedCentral
53.
Gemmill RM, Roche J, Potiron VA, Nasarre P, Mitas M, Coldren CD, Helfrich BA, Garrett-Mayer E, Bunn PA, Drabkin HA: ZEB1-responsive genes in non-small cell lung cancer. Cancer Lett. 2010, 300 (1): 66-78.CrossRefPubMedPubMedCentral
54.
Park SM, Gaur AB, Lengyel E, Peter ME: The miR-200 family determines the epithelial phenotype of cancer cells by targeting the E-cadherin repressors ZEB1 and ZEB2. Genes Dev. 2008, 22 (7): 894-907. 10.1101/gad.1640608.CrossRefPubMedPubMedCentral
55.
Korpal M, Lee ES, Hu G, Kang Y: The miR-200 family inhibits epithelial-mesenchymal transition and cancer cell migration by direct targeting of E-cadherin transcriptional repressors ZEB1 and ZEB2. J Biol Chem. 2008, 283 (22): 14910-14914. 10.1074/jbc.C800074200.CrossRefPubMedPubMedCentral
56.
Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, Vadas MA, Khew-Goodall Y, Goodall GJ: The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol. 2008, 10 (5): 593-601. 10.1038/ncb1722.CrossRefPubMed
Metadata
Title
Identification of genes regulating migration and invasion using a new model of metastatic prostate cancer
Authors
Jacqueline Banyard
Ivy Chung
Matthew Migliozzi
Derek T Phan
Arianne M Wilson
Bruce R Zetter
Diane R Bielenberg
Publication date
01-12-2014
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2014
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-14-387

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