Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Clinical & Experimental Metastasis
Published in: Clinical & Experimental Metastasis 2/2014

01-02-2014 | Research Paper

PITX2 and non-canonical Wnt pathway interaction in metastatic prostate cancer

Published in: Clinical & Experimental Metastasis | Issue 2/2014

Login to get access

Abstract

The non-canonical Wnt pathway, a regulator of cellular motility and morphology, is increasingly implicated in cancer metastasis. In a quantitative PCR array analysis of 84 Wnt pathway associated genes, both non-canonical and canonical pathways were activated in primary and metastatic tumors relative to normal prostate. Expression of the Wnt target gene PITX2 in a prostate cancer (PCa) bone metastasis was strikingly elevated over normal prostate (over 2,000-fold) and primary prostate cancer (over 200-fold). The elevation of PITX2 protein was also evident on tissue microarrays, with strong PITX2 immunostaining in PCa skeletal and, to a lesser degree, soft tissue metastases. PITX2 is associated with cell migration during normal tissue morphogenesis. In our studies, overexpression of individual PITX2A/B/C isoforms stimulated PC-3 PCa cell motility, with the PITX2A isoform imparting a specific motility advantage in the presence of non-canonical Wnt5a stimulation. Furthermore, PITX2 specific shRNA inhibited PC-3 cell migration toward bone cell derived chemoattractant. These experimental results support a pivotal role of PITX2A and non-canonical Wnt signaling in enhancement of PCa cell motility, suggest PITX2 involvement in homing of PCa to the skeleton, and are consistent with a role for PITX2 in PCa metastasis to soft and bone tissues. Our findings, which significantly expand previous evidence that PITX2 is associated with risk of PCa biochemical recurrence, indicate that variation in PITX2 expression accompanies and may promote prostate tumor progression and metastasis.
Appendix
Available only for authorised users
Literature
1.
Hall CL, Kang S, MacDougald OA, Keller ET (2006) Role of wnts in prostate cancer bone metastases. J Cell Biochem 97(4):661–672PubMedCrossRef
2.
Hall CL, Bafico A, Dai J, Aaronson SA, Keller ET (2005) Prostate cancer cells promote osteoblastic bone metastases through Wnts. Cancer Res 65(17):7554–7560PubMed
3.
Glass DA 2nd, Bialek P, Ahn JD, Starbuck M, Patel MS, Clevers H, Taketo MM, Long F, McMahon AP, Lang RA, Karsenty G (2005) Canonical Wnt signaling in differentiated osteoblasts controls osteoclast differentiation [see comment]. Dev Cell 8(5):751–764PubMedCrossRef
4.
5.
Semenov MV, Habas R, MacDonald BT, He X (2007) Snapshot: noncanonical Wnt signaling pathways. Cell 131(7):1378PubMedCrossRef
6.
Veeman MT, Axelrod JD, Moon RT (2003) A second canon: functions and mechanisms of beta-catenin-independent Wnt signaling. Dev Cell 5(3):367–377PubMedCrossRef
7.
8.
9.
Moustakas A, Heldin C-H (2007) Signaling networks guiding epithelial-mesenchymal transitions during embryogenesis and cancer progression. Cancer Sci 98(10):1512–1520PubMedCrossRef
10.
11.
Quentien MH, Barlier A, Franc JL, Pellegrini I, Brue T, Enjalbert A (2006) Pituitary transcription factors: from congenital deficiencies to gene therapy. J Neuroendocrinol 18(9):633–642PubMedCrossRef
12.
Kioussi C, Briata P, Baek SH, Rose DW, Hamblet NS, Herman T, Ohgi KA, Lin CJ, Gleiberman A, Wang JB, Brault V, Ruiz-Lozano P, Nguyen HD, Kemler R, Glass CK, Wynshaw-Boris A, Rosenfeld MG (2002) Identification of a Wnt/DVI/beta-catenin—>Pitx2 pathway mediating cell-type-specific proliferation during development. Cell 111(5):673–685PubMedCrossRef
13.
Amendt BA, Sutherland LB, Russo AF (1999) Multifunctional role of the Pitx2 homeodomain protein C-terminal tail. Mol Cell Biol 19(10):7001–7010PubMedCentralPubMed
14.
Ai D, Liu W, Ma LJ, Dong FY, Lu MF, Wang DG, Verzi MP, Cai CL, Gage PJ, Evans S, Black BL, Brown NA, Martin JF (2006) Pitx2 regulates cardiac left-right asymmetry by patterning second cardiac lineage-derived myocardium. Dev Biol 296(2):437–449PubMedCrossRef
15.
Kurpios NA, Ibanes M, Davis NM, Lui W, Katz T, Martin JF, Belmonte JCI, Tabintt CJ (2008) The direction of gut looping is established by changes in the extracellular matrix and in cell: cell adhesion. Proc Natl Acad Sci USA 105(25):8499–8506PubMedCrossRef
16.
Ai D, Wang J, Amen M, Lu MF, Amendt BA, Martin JF (2007) Nuclear factor 1 and T-cell factor/LEF recognition elements regulate Pitx2 transcription in pituitary development. Mol Cell Biol 27(16):5765–5775PubMedCentralPubMedCrossRef
17.
Amen M, Liu X, Vadlamudi U, Elizondo G, Diamond E, Engelhardt JF, Amendt BA (2007) PITX2 and beta-catenin interactions regulate lef-1 isoform expression. Mol Cell Biol 27(21):7560–7573PubMedCentralPubMedCrossRef
18.
Baek SH, Kioussi C, Briata P, Wang DG, Nguyen HD, Ohgi KA, Glass CK, Wynshaw-Boris A, Rose DW, Rosenfeld MG (2003) Regulated subset of G(1) growth-control genes in response to derepression by the Wnt pathway. Proc Natl Acad Sci USA 100(6):3245–3250PubMedCrossRef
19.
Briata P, Ilengo C, Corte G, Moroni C, Rosenfeld MG, Chen CY, Gherzi R (2003) The Wnt/beta-catenin—>Pitx2 pathway controls the turnover of Pitx2 and other unstable mRNAs. Mol Cell 12(5):1201–1211PubMedCrossRef
20.
Vadlamudi U, Espinoza HM, Ganga M, Martin DM, Liu XM, Engelhardt JF, Amendt BA (2005) PITX2, beta-catenin and LEF-1 interact to synergistically regulate the LEF-1 promoter. J Cell Sci 118(6):1129–1137PubMedCrossRef
21.
Zhou WL, Lin LZ, Majumdar A, Li X, Zhang XX, Liu W, Etheridge L, Shi YQ, Martin J, Van de Ven W, Kaartinen V, Wynshaw-Boris A, McMahon AP, Rosenfeld MG, Evans SM (2007) Modulation of morphogenesis by noncanonical Wnt signaling requires ATF/CREB family-mediated transcriptional activation of TGF beta 2. Nat Genet 39(10):1225–1234PubMedCrossRef
22.
23.
Weiss G, Cottrell S, Distler J, Schatz P, Kristiansen G, Ittmann M, Haefliger C, Lesche R, Hartmann A, Corman J, Wheeler T (2009) DNA methylation of the PITX2 gene promoter region is a strong independent prognostic marker of biochemical recurrence in patients with prostate cancer after radical prostatectomy. J Urol 181(4):1678–1685PubMedCrossRef
24.
Vanaja DK, Ehrich M, Van den Boom D, Cheville JC, Karnes RJ, Tindall DJ, Cantor CR, Young CYF (2009) Hypermethylation of genes for diagnosis and risk stratification of prostate cancer. Cancer Invest 27(5):549–560PubMedCentralPubMedCrossRef
25.
Banez LL, Sun L, van Leenders GJ, Wheeler TM, Bangma CH, Freedland SJ, Ittmann MM, Lark AL, Madden JF, Hartman A, Weiss G, Castanos-Velez E (2010) Multicenter clinical validation of PITX2 methylation as a prostate specific antigen recurrence predictor in patients with post-radical prostatectomy prostate cancer. J Urol 184(1):149–156. doi:10.​1016/​j.​juro.​2010.​03.​012 PubMedCrossRef
26.
Goebel G, Auer D, Gaugg I, Schneitter A, Lesche R, Mueller-Holzner E, Marth C, Daxenbichler G (2011) Prognostic significance of methylated RASSF1A and PITX2 genes in blood- and bone marrow plasma of breast cancer patients. Breast Cancer Res Treat 130(1):109–117. doi:10.​1007/​s10549-010-1335-8 CrossRef
27.
28.
Dietrich D, Lesche R, Tetzner R, Krispin M, Dietrich J, Haedicke W, Schuster M, Kristiansen G (2009) Analysis of DNA methylation of multiple genes in microdissected cells from formalin-fixed and paraffin-embedded Tissues. J Histochem Cytochem 57(5):477–489PubMedCrossRef
29.
Nimmrich I, Sieuwerts AM, Gelder MEM, Schwope I, Vries JB, Harbeck N, Koenig T, Hartmann O, Kluth A, Dietrich D, Magdolen V, Portengen H, Look MP, Klijn JGM, Lesche R, Schmitt M, Maier S, Foekens JA, Martens JWM (2008) DNA hypermethylation of PITX2 is a marker of poor prognosis in untreated lymph node-negative hormone receptor-positive breast cancer patients. Breast Cancer Res Treat 111(3):429–437PubMedCrossRef
30.
Harbeck N, Nimmrich I, Hartmann A, Ross JS, Cufer T, Grutzmann R, Kristiansen G, Paradiso A, Hartmann O, Margossian A, Martens J, Schwope I, Lukas A, Muller V, Milde-Langosch K, Nahrig J, Foekens J, Maier S, Schmitt M, Lesche R (2008) Multicenter study using paraffin-embedded tumor tissue testing Pitx2 DNA methylation as a marker for outcome prediction in tamoxifen-treated, node-negative breast cancer patients. J Clin Oncol 26(31):5036–5042. doi:10.​1200/​jco.​2007.​14.​1697 PubMedCrossRef
31.
Span PN, Sieuwerts AM, Heuvel J, Spyratos F, Duffy MJ, Eppenberger-Castori S, Vacher S, O’Brien K, McKiernan E, Pierce A, Vuaroqueaux V, Foekens JA, Sweep F, Martens JWM (2009) Harmonisation of multi-centre real-time reverse-transcribed PCR results of a candidate prognostic marker in breast cancer: an EU-FP6 supported study of members of the EORTC—pathobiology group. Eur J Cancer 45(1):74–81PubMedCrossRef
32.
Roudier MP, True LD, Higano CS, Vesselle H, Ellis W, Lange P, Vessella RL (2003) Phenotypic heterogeneity of end-stage prostate carcinoma metastatic to bone. Hum Pathol 34(7):646–653. doi:S004681770300190​4 [pii]PubMedCrossRef
33.
34.
Lamba P, Hjalt T, Bernard D (2008) Novel forms of paired-like homeodomain transcription factor 2 (PITX2): generation by alternative translation initiation and mRNA splicing. BMC Mol Biol 9(1):31PubMedCentralPubMedCrossRef
35.
Saito K, Oku T, Ata N, Miyashiro H, Hattori M, Saiki I (1997) A modified and convenient method for assessing tumor cell invasion and migration and its application to screening for inhibitors. Biol Pharm Bull 20(4):345–348PubMedCrossRef
36.
Rosen S, Skaletsky HJ (2000) Primer 3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols: methods in molecular biology. Humana Press, Totowa, pp 365–386
37.
Wei Q, Adelstein RS (2002) Pitx2a expression alters actin-myosin cytoskeleton and migration of HeLa cells through Rho GTPase signaling. Mol Biol Cell 13(2):683–697PubMedCentralPubMedCrossRef
38.
Gage PJ, Qian M, Wu DQ, Rosenberg KI (2008) The canonical Wnt signaling antagonist DKK2 is an essential effector of PITX2 function during normal eye development. Dev Biol 317(1):310–324PubMedCentralPubMedCrossRef
39.
Vinarskaja A, Schulz WA, Ingenwerth M, Hader C, Arsov C (2011) Association of PITX2 mRNA down-regulation in prostate cancer with promoter hypermethylation and poor prognosis. Urol Oncol 31(5):621–627. doi:10.​1016/​j.​urolonc.​2011.​04.​010 S1078-1439(11)00139-6 [pii]
40.
Shiratori H, Sakuma R, Watanabe M, Hashiguchi H, Mochida K, Sakai Y, Nishino J, Saijoh Y, Whitman M, Hamada H (2001) Two-step regulation of left–right asymmetric expression of Pitx2: initiation by nodal signaling and maintenance by Nkx2. Mol Cell 7(1):137–149. doi:S1097-2765(01)00162-9 [pii]PubMedCrossRef
41.
42.
Shima Y, Zubair M, Komatsu T, Oka S, Yokoyama C, Tachibana T, Hjalt TA, Drouin J, Morohashi K (2008) Pituitary homeobox 2 regulates adrenal4 binding protein/steroidogenic factor-1 gene transcription in the pituitary gonadotrope through interaction with the intronic enhancer. Mol Endocrinol 22(7):1633–1646PubMedCrossRef
43.
Hall CL, Dubyk CW, Riesenberger TA, Shein D, Keller ET, van Golen K (2008) Type I collagen receptor (alpha(2)beta(1)) signaling promotes prostate cancer invasion through RhoC GTPase. Neoplasia 10(8):797–803PubMedCentralPubMed
44.
Hayashi M, Maeda S, Aburatani H, Kitamura K, Miyoshi H, Miyazono K, Imamura T (2008) Pitx2 prevents osteoblastic transdifferentiation of myoblasts by bone morphogenetic proteins. J Biol Chem 283(1):565–571. doi:10.​1074/​jbc.​M708154200 PubMedCrossRef
45.
Zhu L, Marvin MJ, Gardiner A, Lassar AB, Mercola M, Stern CD, Levin M (1999) Cerberus regulates left–right asymmetry of the embryonic head and heart. Curr Biol 9(17):931–938PubMedCrossRef
46.
Suszko MI, Antenos M, Balkin DM, Woodruff TK (2008) Smad3 and Pitx2 cooperate in stimulation of FSH beta gene transcription. Mol Cell Endocrinol 281(1–2):27–36PubMedCrossRef
47.
48.
Basu M, Roy SS (2012) Wnt/β-catenin pathway is regulated by PITX2 homeodomain protein and thus contributes to the proliferation of human ovarian adenocarcinoma cell, SKOV-3. J Biol Chem. doi:10.​1074/​jbc.​M112.​409102
49.
50.
Huang Y, Guigon CJ, Fan J, Cheng S-Y, Zhu G-Z (2010) Pituitary homeobox 2 (PITX2) promotes thyroid carcinogenesis by activation of cyclin D2. Cell Cycle 9(7):1333–1341PubMedCrossRef
51.
Aitchison AA, Veerakumarasivam A, Vias M, Kumar R, Hamdy FC, Neal DE, Milis IG (2008) Promoter methylation correlates with reduced Smad4 expression in advanced prostate cancer. Prostate 68(6):661–674PubMedCrossRef
52.
Connolly RM, Visvanathan K (2008) PITX2 DNA methylation: a potential prognostic or predictive biomarker in early breast cancer. Pharmacogenomics 9(12):1797–1798CrossRef
53.
Connolly RM, Visvanathan K (2008) PITX2 DNA methylation as a marker for outcome prediction in breast cancer patients receiving adjuvant tamoxifen. Pharmacogenomics 9(12):1798–1799CrossRef
54.
Maier S, Nimmrich I, Koenig T, Eppenberger-Castori S, Bohlmann I, Paradiso A, Spyratos F, Thomssen C, Mueller V, Naehrig J, Schittulli F, Kates R, Lesche R, Schwope I, Kluth A, Marx A, Martens JWM, Foekens JA, Schmitt M, Harbeck N (2007) DNA-methylation of the homeodomain transcription factor PITX2 reliably predicts risk of distant disease recurrence in tamoxifen-treated, node-negative breast cancer patients Technical and clinical validation in a multi-centre setting in collaboration with the European Organisation for Research and Treatment of Cancer (EORTC) PathoBiology group. Eur J Cancer 43(11):1679–1686PubMedCrossRef
55.
56.
Metadata
Title
PITX2 and non-canonical Wnt pathway interaction in metastatic prostate cancer
Publication date
01-02-2014
Published in
Clinical & Experimental Metastasis / Issue 2/2014
Print ISSN: 0262-0898
Electronic ISSN: 1573-7276
DOI
https://doi.org/10.1007/s10585-013-9620-7

Other articles of this Issue 2/2014

Clinical & Experimental Metastasis 2/2014 Go to the issue

Research Paper

Primary 4T1 tumor resection provides critical “window of opportunity” for immunotherapy

Research Paper

RANK expression on breast cancer cells promotes skeletal metastasis

Research Paper

Percutaneous image-guided biopsy of prostate cancer metastases yields samples suitable for genomics and personalised oncology

Review Paper

Breaking and entering into the CNS: clues from solid tumor and nonmalignant models with relevance to hematopoietic malignancies

Research Paper

Morphine and breast tumor metastasis: the role of matrix-degrading enzymes

Research Paper

Changes in the peripheral blood and bone marrow from untreated advanced breast cancer patients that are associated with the establishment of bone metastases
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits