Journal Article
. 2014 Dec; 20(46):17376-87.
doi: 10.3748/wjg.v20.i46.17376.

Pea3 expression promotes the invasive and metastatic potential of colorectal carcinoma

Aruz Mesci 1 Samira Taeb 1 Xiaoyong Huang 1 Rishi Jairath 1 Darshan Sivaloganathan 1 Stanley K Liu 1 
  • PMID: 25516649
  •     43 References
  •     12 citations


Aim: To investigate the function of Pea3 in colorectal carcinoma (CRC) invasion and metastatic potential.

Methods: The expression of Pea3 during clinical progression of human CRC was investigated using Oncomine Research Edition. To assay Pea3 expression in established CRC cell lines, we performed western blotting of cell lysates. We employed shRNA-mediated knockdown of Pea3 in HCT116 (HCT) and LS174T CRC cells which was confirmed by real-time quantitative PCR (qPCR) and western blotting. Transwell invasion assays, MTS proliferation assays, anoikis assays, and fluorometric matrix metalloprotease (MMP) assays were performed to determine the effects of Pea3 knockdown on invasion, proliferation, anoikis and MMP activity in CRC cells in vitro. Alterations in epithelial-mesenchymal transition (EMT) and matrix metalloprotease (MMP) mRNA levels were determined by qPCR. CRC cells were injected into the flanks of nude mice to generate xenografts and tumor growth monitored with serial calliper measurements. To assay metastatic potential, CRC cells were injected into the spleen of nude mice, and histological analysis performed on the livers 21 d later.

Results: We demonstrated that reduction of Pea3 expression in CRC cells significantly impaired their invasive capacity (HCT.shPea3, 0.28 ± 0.04 fold, P < 0.01; LS.shPea3, 0.15 ± 0.04 fold; SW.shPea3, 0.23 ± 0.03, P < 0.01), reduced anoikis resistance (HCT.shPea3 75.4% ± 1.9% viable cells vs HCT.shCtrl 88.6% ± 0.6% viable cells, P < 0.01; LS.shPea3 71.7% ± 0.5% viable cells vs LS.Ctrl 89.6% ± 0.3% viable cells, P < 0.005, but had no effect on proliferation (HCT.shCtrl AUC 5098 ± 123 vs HCT.shPea3 5689 ± 151, P < 0.05; LS.shCtrl AUC 5600 ± 324.1 vs LS.shPea3 6423 ± 400, P < 0.05). In vivo, HCT.shPea3 and HCT.shCtrl tumour xenografts grew at a similar rate (HCT.shPea3 2.64 ± 0.82 fold vs HCT.shCtrl 2.88 ± 0.80 fold, P > 0.05). In keeping with a pro-metastatic function for Pea3 in CRC, several EMT markers and MMPs were downregulated in shPea3-expressing cells, suggesting that Pea3 may exert its effects through these processes. A reduction in overall MMP activity was observed in HCT.shPea3 cells compared to their control counterparts (HCT.shPea3 0.61 ± 0.04 fold, P < 0.005). This translated in vivo to the complete absence of metastases in the livers of mice that were grafted with CRC cells lacking Pea3. Conversely, CRC cells expressing Pea3 formed liver metastases in all mice.

Conclusion: Our study implicates Pea3 as a mediator of metastases, and provides a biological rationale for the adverse prognosis associated with elevated Pea3 expression in human CRC.

Keywords: Colorectal cancer; Epithelial-mesenchymal transition; Liver metastasis; Pea3.

The PEA3 group of ETS-related transcription factors. Role in breast cancer metastasis.
Y de Launoit, A Chotteau-Lelievre, +5 authors, J L Baert.
Adv Exp Med Biol, 2000 Aug 26; 480. PMID: 10959416
ETV4 promotes metastasis in response to activation of PI3-kinase and Ras signaling in a mouse model of advanced prostate cancer.
Alvaro Aytes, Antonina Mitrofanova, +9 authors, Cory Abate-Shen.
Proc Natl Acad Sci U S A, 2013 Aug 07; 110(37). PMID: 23918374    Free PMC article.
PEA3 transcription factors are expressed in tissues undergoing branching morphogenesis and promote formation of duct-like structures by mammary epithelial cells in vitro.
Anne Chotteau-Lelievre, Roberto Montesano, +3 authors, Yvan de Launoit.
Dev Biol, 2003 Jul 23; 259(2). PMID: 12871699
Association of ets-related transcriptional factor E1AF expression with tumour progression and overexpression of MMP-1 and matrilysin in human colorectal cancer.
Shina Horiuchi, Hiroyuki Yamamoto, +3 authors, Kohzoh Imai.
J Pathol, 2003 Aug 05; 200(5). PMID: 12898592
Expression of PEA3/E1AF/ETV4, an Ets-related transcription factor, in breast tumors: positive links to MMP2, NRG1 and CGB expression.
Ivan Bièche, Sengül Tozlu, +4 authors, Rosette Lidereau.
Carcinogenesis, 2003 Nov 25; 25(3). PMID: 14633660
Role of matrix metalloproteinases (MMPs) in colorectal cancer.
Stanley Zucker, Jeffrey Vacirca.
Cancer Metastasis Rev, 2004 Mar 06; 23(1-2). PMID: 15000152
Highly Cited. Review.
Polyoma enhancer activator 3, an ets transcription factor, mediates the induction of cyclooxygenase-2 by nitric oxide in colorectal cancer cells.
Yongmin Liu, Gregory L Borchert, James M Phang.
J Biol Chem, 2004 Feb 21; 279(18). PMID: 14976201
Epidermal growth factor upregulates matrix metalloproteinase-7 expression through activation of PEA3 transcription factors.
Conor C Lynch, Howard C Crawford, Lynn M Matrisian, Susan McDonnell.
Int J Oncol, 2004 May 13; 24(6). PMID: 15138601
Expression of SRC-1, AIB1, and PEA3 in HER2 mediated endocrine resistant breast cancer; a predictive role for SRC-1.
F J Fleming, E Myers, +5 authors, L S Young.
J Clin Pathol, 2004 Sep 29; 57(10). PMID: 15452162    Free PMC article.
E1AF, an ets-oncogene family transcription factor.
Masanobu Shindoh, Fumihiro Higashino, Takao Kohgo.
Cancer Lett, 2004 Oct 27; 216(1). PMID: 15500943
A new animal model for human colon cancer metastasis.
R S Bresalier, S E Raper, E S Hujanen, Y S Kim.
Int J Cancer, 1987 May 15; 39(5). PMID: 3032811
Isolation of a cDNA encoding the adenovirus E1A enhancer binding protein: a new human member of the ets oncogene family.
F Higashino, K Yoshida, +2 authors, K Fujinaga.
Nucleic Acids Res, 1993 Feb 11; 21(3). PMID: 8441666    Free PMC article.
Stimulation of 92-kDa gelatinase B promoter activity by ras is mitogen-activated protein kinase kinase 1-independent and requires multiple transcription factor binding sites including closely spaced PEA3/ets and AP-1 sequences.
R Gum, E Lengyel, +4 authors, D Boyd.
J Biol Chem, 1996 May 03; 271(18). PMID: 8631874
Differential expression patterns of the PEA3 group transcription factors through murine embryonic development.
A Chotteau-Lelièvre, X Desbiens, +2 authors, Y de Launoit.
Oncogene, 1997 Aug 18; 15(8). PMID: 9285689
HER2/Neu and the Ets transcription activator PEA3 are coordinately upregulated in human breast cancer.
C C Benz, R C O'Hagan, +8 authors, J A Hassell.
Oncogene, 1997 Oct 31; 15(13). PMID: 9380403
Increased E1AF expression in mouse fibrosarcoma promotes metastasis through induction of MT1-MMP expression.
H Habelhah, F Okada, +8 authors, M Hosokawa.
Oncogene, 1999 Apr 20; 18(9). PMID: 10208438
Association of Ets-related transcriptional factor E1AF expression with overexpression of matrix metalloproteinases, COX-2 and iNOS in the early stage of colorectal carcinogenesis.
Katsuhiko Nosho, Mio Yoshida, +5 authors, Kohzoh Imai.
Carcinogenesis, 2005 Feb 08; 26(5). PMID: 15695237
E1A-F is overexpressed early in human colorectal neoplasia and associated with cyclooxygenase-2 and matrix metalloproteinase-7.
William M Boedefeld, Richie Soong, +4 authors, Martin J Heslin.
Mol Carcinog, 2005 Apr 01; 43(1). PMID: 15800927
E1AF/PEA3 activates the Rho/Rho-associated kinase pathway to increase the malignancy potential of non-small-cell lung cancer cells.
Nobuyuki Hakuma, Ichiro Kinoshita, +4 authors, Hirotoshi Dosaka-Akita.
Cancer Res, 2005 Dec 03; 65(23). PMID: 16322223
Tumor invasion: role of growth factor-induced cell motility.
A Wells.
Adv Cancer Res, 1999 Nov 05; 78. PMID: 10547668
A positive role for PEA3 in HER2-mediated breast tumour progression.
E Myers, A D K Hill, +3 authors, L S Young.
Br J Cancer, 2006 Oct 25; 95(10). PMID: 17060941    Free PMC article.
Development and characterization of a model of liver metastasis using human colon cancer HCT-116 cells.
Kazuhiro Ishizu, Naohide Sunose, +4 authors, Takao Yamori.
Biol Pharm Bull, 2007 Sep 11; 30(9). PMID: 17827739
Invasion assays.
H K Kleinman, K Jacob.
Curr Protoc Cell Biol, 2008 Jan 30; Chapter 12. PMID: 18228316
FGF-regulated Etv genes are essential for repressing Shh expression in mouse limb buds.
Zhen Zhang, Jamie M Verheyden, John A Hassell, Xin Sun.
Dev Cell, 2009 Apr 24; 16(4). PMID: 19386269    Free PMC article.
Increased PEA3/E1AF and decreased Net/Elk-3, both ETS proteins, characterize human NSCLC progression and regulate caveolin-1 transcription in Calu-1 and NCI-H23 NSCLC cell lines.
Karin A Sloan, Hector A Marquez, +7 authors, Hasmeena Kathuria.
Carcinogenesis, 2009 Jun 02; 30(8). PMID: 19483189    Free PMC article.
S100A4 and metastasis: a small actor playing many roles.
Kjetil Boye, Gunhild M Maelandsmo.
Am J Pathol, 2009 Dec 19; 176(2). PMID: 20019188    Free PMC article.
Highly Cited. Review.
Osteopontin provides early proliferative drive and may be dependent upon aberrant c-myc signalling in murine intestinal tumours.
Cristina Martinez, Michael Churchman, Tom Freeman, Mohammad Ilyas.
Exp Mol Pathol, 2010 Jan 08; 88(2). PMID: 20053348
The ERK MAP kinase-PEA3/ETV4-MMP-1 axis is operative in oesophageal adenocarcinoma.
Richard Keld, Baoqiang Guo, +3 authors, Andrew D Sharrocks.
Mol Cancer, 2010 Dec 15; 9. PMID: 21143918    Free PMC article.
Expression of osteopontin coregulators in primary colorectal cancer and associated liver metastases.
D J Mole, C O'Neill, +6 authors, F C Campbell.
Br J Cancer, 2011 Feb 24; 104(6). PMID: 21343932    Free PMC article.
Polyomavirus enhancer activator 3 protein promotes breast cancer metastatic progression through Snail-induced epithelial-mesenchymal transition.
Hiu-Fung Yuen, Yuen-Kwong Chan, +10 authors, Mohamed El-Tanani.
J Pathol, 2011 Mar 16; 224(1). PMID: 21404275
PEA3/ETV4-related transcription factors coupled with active ERK signalling are associated with poor prognosis in gastric adenocarcinoma.
R Keld, B Guo, +4 authors, A D Sharrocks.
Br J Cancer, 2011 Jun 16; 105(1). PMID: 21673681    Free PMC article.
COX-2 gene expression in colon cancer tissue related to regulating factors and promoter methylation status.
Annika Gustafsson Asting, Helena Carén, +3 authors, Kent Lundholm.
BMC Cancer, 2011 Jun 15; 11. PMID: 21668942    Free PMC article.
Current progress of siRNA/shRNA therapeutics in clinical trials.
John C Burnett, John J Rossi, Katrin Tiemann.
Biotechnol J, 2011 Jul 12; 6(9). PMID: 21744502    Free PMC article.
Highly Cited. Review.
Anoikis: an emerging hallmark in health and diseases.
M L Taddei, E Giannoni, T Fiaschi, P Chiarugi.
J Pathol, 2011 Sep 29; 226(2). PMID: 21953325
Highly Cited. Review.
Delta-like ligand 4-notch blockade and tumor radiation response.
Stanley K Liu, Saif A S Bham, +5 authors, Ruth J Muschel.
J Natl Cancer Inst, 2011 Oct 20; 103(23). PMID: 22010178
Knockdown of S100A4 decreases tumorigenesis and metastasis in osteosarcoma cells by repression of matrix metalloproteinase-9.
Guoyou Zhang, Ming Li, +2 authors, Changwei Yang.
Asian Pac J Cancer Prev, 2012 Feb 02; 12(8). PMID: 22292654
ETV1, 4 and 5: an oncogenic subfamily of ETS transcription factors.
Sangphil Oh, Sook Shin, Ralf Janknecht.
Biochim Biophys Acta, 2012 Mar 20; 1826(1). PMID: 22425584    Free PMC article.
Highly Cited. Review.
Epithelial mesenchymal transition in colorectal cancer: Seminal role in promoting disease progression and resistance to neoadjuvant therapy.
Aneel Bhangu, Gemma Wood, +3 authors, Robert Goldin.
Surg Oncol, 2012 Sep 18; 21(4). PMID: 22981546
S100A4 promotes invasion and angiogenesis in breast cancer MDA-MB-231 cells by upregulating matrix metalloproteinase-13.
Lin Wang, Xingang Wang, +2 authors, Qingfeng Chen.
Acta Biochim Pol, 2012 Nov 20; 59(4). PMID: 23162804
Pathways of colorectal carcinogenesis.
Mark J Arends.
Appl Immunohistochem Mol Morphol, 2013 Feb 19; 21(2). PMID: 23417071
Targeted therapy in metastatic colorectal cancer -- an example of personalised medicine in action.
V Heinemann, J Y Douillard, M Ducreux, M Peeters.
Cancer Treat Rev, 2013 Feb 05; 39(6). PMID: 23375249
S100A4 silencing suppresses proliferation, angiogenesis and invasion of thyroid cancer cells through downregulation of MMP-9 and VEGF.
W Jia, X-J Gao, +2 authors, G Zhang.
Eur Rev Med Pharmacol Sci, 2013 Jun 19; 17(11). PMID: 23771538
Tumor invasion as dysregulated cell motility.
J Kassis, D A Lauffenburger, T Turner, A Wells.
Semin Cancer Biol, 2001 Apr 27; 11(2). PMID: 11322830
PEA3 transcription factors are downstream effectors of Met signaling involved in migration and invasiveness of Met-addicted tumor cells.
Zoulika Kherrouche, Didier Monte, +5 authors, Anne Chotteau-Lelievre.
Mol Oncol, 2015 Aug 05; 9(9). PMID: 26238631    Free PMC article.
Targeting of CCBE1 by miR-330-3p in human breast cancer promotes metastasis.
Aruz Mesci, Xiaoyong Huang, +6 authors, Stanley K Liu.
Br J Cancer, 2017 Apr 19; 116(10). PMID: 28419078    Free PMC article.
ETV4 Facilitates Cell-Cycle Progression in Pancreatic Cells through Transcriptional Regulation of Cyclin D1.
Nikhil Tyagi, Sachin K Deshmukh, +7 authors, Seema Singh.
Mol Cancer Res, 2017 Nov 10; 16(2). PMID: 29117940    Free PMC article.
ETV4 transcription factor and MMP13 metalloprotease are interplaying actors of breast tumorigenesis.
Mandy Dumortier, Franck Ladam, +6 authors, Anne Chotteau-Lelièvre.
Breast Cancer Res, 2018 Jul 13; 20(1). PMID: 29996935    Free PMC article.
The Transcription Factor ETV5 Mediates BRAFV600E-Induced Proliferation and TWIST1 Expression in Papillary Thyroid Cancer Cells.
Oorvashi Roy Puli, Brian P Danysh, +7 authors, Marie-Claude Hofmann.
Neoplasia, 2018 Sep 29; 20(11). PMID: 30265861    Free PMC article.
Capicua suppresses colorectal cancer progression via repression of ETV4 expression.
Jeon-Soo Lee, Eunjeong Kim, +6 authors, Yoontae Lee.
Cancer Cell Int, 2020 Feb 12; 20. PMID: 32042269    Free PMC article.
Function and regulation of the PEA3 subfamily of ETS transcription factors in cancer.
Tingting Qi, Qiang Qu, +6 authors, Jian Qu.
Am J Cancer Res, 2020 Nov 10; 10(10). PMID: 33163259    Free PMC article.
ACE2 Is a Prognostic Biomarker and Associated with Immune Infiltration in Kidney Renal Clear Cell Carcinoma: Implication for COVID-19.
Xinhao Niu, Zhe Zhu, Enming Shao, Juan Bao.
J Oncol, 2021 Feb 11; 2021. PMID: 33564310    Free PMC article.
The E-Twenty-Six Family in Hepatocellular Carcinoma: Moving into the Spotlight.
Tongyue Zhang, Danfei Liu, +2 authors, Limin Xia.
Front Oncol, 2021 Feb 16; 10. PMID: 33585247    Free PMC article.
ETV4 plays a role on the primary events during the adenoma-adenocarcinoma progression in colorectal cancer.
Aline Simoneti Fonseca, Anelisa Ramão, +17 authors, Wilson Araújo Silva.
BMC Cancer, 2021 Mar 03; 21(1). PMID: 33648461    Free PMC article.
Gene Regulatory Network of ETS Domain Transcription Factors in Different Stages of Glioma.
Yigit Koray Babal, Basak Kandemir, Isil Aksan Kurnaz.
J Pers Med, 2021 Mar 07; 11(2). PMID: 33671331    Free PMC article.
Dysregulation of vitamin D synthesis pathway genes in colorectal cancer: A case-control study.
Hossein Sadeghi, Zeeba Kamaliyan, +7 authors, Arfa Moshiri.
J Clin Lab Anal, 2020 Oct 16; 35(2). PMID: 33058307    Free PMC article.