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BMC Cancer
Published in: BMC Cancer 1/2016

Open Access 01-12-2016 | Research article

ROR2 is epigenetically inactivated in the early stages of colorectal neoplasia and is associated with proliferation and migration

Authors: Sean S. Q. Ma, Sameer Srivastava, Estelle Llamosas, Nicholas J. Hawkins, Luke B. Hesson, Robyn L. Ward, Caroline E. Ford

Published in: BMC Cancer | Issue 1/2016

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Abstract

Background

Colorectal cancer (CRC) is closely linked to Wnt signalling, with 94 % of cases exhibiting a Wnt related mutation. ROR2 is a receptor tyrosine kinase that is thought to repress β-catenin dependent Wnt signalling. Our study aims to determine if ROR2 is epigenetically silenced in CRC and determine if in vitro silencing of ROR2 potentiates Wnt signalling, and alters the proliferative, migratory or invasive potential of cells.

Methods

ROR2 expression was examined in CRC cell lines and patient adenomas using qRT-PCR, while COBRA and bisulphite sequencing was used to analyse ROR2 promoter methylation. 258 patient primary tumour samples from publicly available databases were also examined for ROR2 expression and methylation. In addition, the functional effects of ROR2 modulation were investigated in HCT116 cells following ROR2 siRNA knockdown and in RKO and SW620 cells following ectopic ROR2 expression.

Results

Reduced ROR2 expression was found to correlate with ROR2 promoter hypermethylation in colorectal cancer cell lines, carcinomas and adenomas. ROR2 expression was downregulated in 76.7 % (23/30) of CRC cell lines with increasing ROR2 promoter hypermethylation correlating with progressively lower expression. Analysis of 239 primary tumour samples from a publicly available cohort also found a significant correlation between reduced ROR2 expression and increased promoter methylation. Methylation analysis of 88 adenomas and 47 normal mucosa samples found greater percentage of adenoma samples to be methylated. Additional analysis also revealed that adenoma samples with reduced ROR2 expression also possessed ROR2 promoter hypermethylation. ROR2 knockdown in the CRC cell line HCT116 significantly decreased expression of the β-catenin independent Wnt targets genes JNK and NFATC1, increased cellular proliferation and migration but decreased invasion. When ROR2 was ectopically expressed in RKO and SW620 cells, there was no significant change to either cellular proliferation or migration.

Conclusion

ROR2 is frequently epigenetically inactivated by promoter hypermethylation in the early stages of colorectal neoplasia and this may contribute to colorectal cancer progression by increasing cellular proliferation and migration.
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Literature
1.
Ferlay J, et al. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 2010;127(12):2893–917.CrossRefPubMed
2.
3.
Siegel R, Desantis C, Jemal A. Colorectal cancer statistics. CA Cancer J Clin. 2014;64(2):104–17.CrossRefPubMed
4.
5.
Markowitz SD, Bertagnolli MM. Molecular Basis of Colorectal Cancer REPLY. N Engl J Med. 2010;362(13):1246–7.
6.
Cancer Genome Atlas, N. Comprehensive molecular characterization of human colon and rectal cancer. Nature. 2012;487(7407):330–7.CrossRef
7.
Fodde R, Smits R, Clevers H. APC, signal transduction and genetic instability in colorectal cancer. Nat Rev Cancer. 2001;1(1):55–67.CrossRefPubMed
8.
Rowan AJ, et al. APC mutations in sporadic colorectal tumors: A mutational “hotspot” and interdependence of the “two hits”. Proc Natl Acad Sci U S A. 2000;97(7):3352–7.CrossRefPubMedPubMedCentral
9.
Ichii S, et al. Inactivation of both APC alleles in an early stage of colon adenomas in a patient with familial adenomatous polyposis (FAP). Hum Mol Genet. 1992;1(6):387–90.CrossRefPubMed
10.
11.
Aguilera O, et al. Epigenetic alterations of the Wnt/beta-catenin pathway in human disease. Endocr Metab Immune Disord Drug Targets. 2007;7(1):13–21.CrossRefPubMed
12.
13.
Voronkov A, Krauss S. Wnt/beta-catenin signaling and small molecule inhibitors. Curr Pharm Des. 2013;19(4):634–64.CrossRefPubMed
14.
Rubinfeld B, et al. Association of the APC gene product with beta-catenin. Science. 1993;262(5140):1731–4.CrossRefPubMed
15.
16.
Oishi I, et al. The receptor tyrosine kinase Ror2 is involved in non-canonical Wnt5a/JNK signalling pathway. Genes Cells. 2003;8(7):645–54.CrossRefPubMed
17.
Katoh M. WNT/PCP signaling pathway and human cancer (review). Oncol Rep. 2005;14(6):1583–8.PubMed
18.
Katoh M, Katoh M. WNT signaling pathway and stem cell signaling network. Clin Cancer Res. 2007;13(14):4042–5.CrossRefPubMed
19.
20.
Debebe Z, Rathmell WK. Ror2 as a Therapeutic Target in Cancer. Pharmacol Ther. 2015.
21.
Matsuda T, et al. Expression of the receptor tyrosine kinase genes, Ror1 and Ror2, during mouse development. Mech Dev. 2001;105(1–2):153–6.CrossRefPubMed
22.
Afzal AR, Jeffery S. One gene, two phenotypes: ROR2 mutations in autosomal recessive Robinow syndrome and autosomal dominant brachydactyly type B. Hum Mutat. 2003;22(1):1–11.CrossRefPubMed
23.
24.
Mikels AJ, Nusse R. Purified Wnt5a protein activates or inhibits beta-catenin-TCF signaling depending on receptor context. Plos Biology. 2006;4(4):570–82.CrossRef
25.
Yamamoto H, et al. Wnt5a modulates glycogen synthase kinase 3 to induce phosphorylation of receptor tyrosine kinase Ror2. Genes Cells. 2007;12(11):1215–23.CrossRefPubMed
26.
Nomachi A, et al. Receptor tyrosine kinase Ror2 mediates Wnt5a-induced polarized cell migration by activating c-Jun N-terminal kinase via actin-binding protein filamin A. J Biol Chem. 2008;283(41):27973–81.CrossRefPubMed
27.
28.
Mei H, et al. High expression of ROR2 in cancer cell correlates with unfavorable prognosis in colorectal cancer. Biochem Biophys Res Commun. 2014;453:703.CrossRefPubMed
29.
O’Connell MP, et al. The orphan tyrosine kinase receptor, ROR2, mediates Wnt5A signaling in metastatic melanoma. Oncogene. 2010;29(1):34–44.CrossRefPubMed
30.
Morioka K, et al. Orphan receptor tyrosine kinase ROR2 as a potential therapeutic target for osteosarcoma. Cancer Sci. 2009;100(7):1227–33.CrossRefPubMed
31.
Enomoto M, et al. Autonomous regulation of osteosarcoma cell invasiveness by Wnt5a/Ror2 signaling. Oncogene. 2009;28(36):3197–208.CrossRefPubMed
32.
Lu BJ, et al. Expression of WNT-5a and ROR2 correlates with disease severity in osteosarcoma. Mol Med Rep. 2012;5(4):1033–6.PubMedPubMedCentral
33.
Henry C, et al. Expression of the novel Wnt receptor ROR2 is increased in breast cancer and may regulate both beta-catenin dependent and independent Wnt signalling. J Cancer Res Clin Oncol. 2015;141(2):243–54.CrossRefPubMed
34.
Ford CE, et al. The dual role of the novel Wnt receptor tyrosine kinase, ROR2, in human carcinogenesis. Int J Cancer. 2012;133(4):779–87.CrossRef
35.
Rasmussen NR, et al. Receptor tyrosine kinase-like orphan receptor 2 (Ror2) expression creates a poised state of Wnt signaling in renal cancer. J Biol Chem. 2013;288(36):26301–10.CrossRefPubMedPubMedCentral
36.
Katoh M, Katoh M. Comparative genomics on ROR1 and ROR2 orthologs. Oncol Rep. 2005;14(5):1381–4.PubMed
37.
Ueno K, et al. Down-regulation of frizzled-7 expression decreases survival, invasion and metastatic capabilities of colon cancer cells. Br J Cancer. 2009;101(8):1374–81.CrossRefPubMedPubMedCentral
38.
Nishita M, et al. Ror2/Frizzled complex mediates Wnt5a-induced AP-1 activation by regulating Dishevelled polymerization. Mol Cell Biol. 2010;30(14):3610–9.CrossRefPubMedPubMedCentral
39.
Asad M, et al. FZD7 drives in vitro aggressiveness in Stem-A subtype of ovarian cancer via regulation of non-canonical Wnt/PCP pathway. Cell Death Dis. 2014;5, e1346.CrossRefPubMedPubMedCentral
40.
Liu Y, et al. Wnt5a induces homodimerization and activation of Ror2 receptor tyrosine kinase. J Cell Biochem. 2008;105(2):497–502.CrossRefPubMed
41.
Kani S, et al. The receptor tyrosine kinase Ror2 associates with and is activated by casein kinase Iepsilon. J Biol Chem. 2004;279(48):50102–9.CrossRefPubMed
42.
Rios AC, et al. Neural crest regulates myogenesis through the transient activation of NOTCH. Nature. 2011;473(7348):532–5.CrossRefPubMed
43.
44.
Faux MC, et al. Restoration of full-length adenomatous polyposis coli (APC) protein in a colon cancer cell line enhances cell adhesion. J Cell Sci. 2004;117(Pt 3):427–39.PubMed
45.
Shan J, et al. Synthesis of potent dishevelled PDZ domain inhibitors guided by virtual screening and NMR studies. Chem Biol Drug Des. 2012;79(4):376–83.CrossRefPubMedPubMedCentral
46.
Madeira M, et al. Estrogen receptor alpha/beta ratio and estrogen receptor beta as predictors of endocrine therapy responsiveness-a randomized neoadjuvant trial comparison between anastrozole and tamoxifen for the treatment of postmenopausal breast cancer. BMC Cancer. 2013;13:425.CrossRefPubMedPubMedCentral
47.
Mitchell RJ, et al. Mismatch repair genes hMLH1 and hMSH2 and colorectal cancer: a HuGE review. Am J Epidemiol. 2002;156(10):885–902.CrossRefPubMed
48.
49.
50.
Hewitt RE, et al. Validation of a model of colon cancer progression. J Pathol. 2000;192(4):446–54.CrossRefPubMed
51.
Sato T, et al. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium. Gastroenterology. 2011;141(5):1762–72.CrossRefPubMed
52.
Ren D, Minami Y, Nishita M. Critical role of Wnt5a-Ror2 signaling in motility and invasiveness of carcinoma cells following Snail-mediated epithelial-mesenchymal transition. Genes Cells. 2011;16(3):304–15.CrossRefPubMed
53.
Li X, et al. Activation of Wnt5a-Ror2 signaling associated with epithelial-to-mesenchymal transition of tubular epithelial cells during renal fibrosis. Genes Cells. 2013;18(7):608–19.CrossRefPubMed
54.
Metz AJ, et al. A correlation of the endoscopic characteristics of colonic laterally spreading tumours with genetic alterations. Eur J Gastroenterol Hepatol. 2013;25(3):319–26.CrossRefPubMed
55.
Metadata
Title
ROR2 is epigenetically inactivated in the early stages of colorectal neoplasia and is associated with proliferation and migration
Authors
Sean S. Q. Ma
Sameer Srivastava
Estelle Llamosas
Nicholas J. Hawkins
Luke B. Hesson
Robyn L. Ward
Caroline E. Ford
Publication date
01-12-2016
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2016
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/s12885-016-2576-7

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