Abstract
Erythropoietin-producing hepatocyte (Eph) receptor family constitutes the largest family of tyrosine kinase receptors in the human genome. Loss of EphB6, a kinase-deficient receptor, correlated with a negative outcome in several carcinomas. This study aimed to investigate the expression of EphB6 protein and mRNA levels in colorectal cancers (CRCs) and possible correlations with clinicopathological variables and prognosis. To assess protein expression level, 124 CRCs and 57 colorectal adenomas samples were examined by immunostaining, the mRNA level of 43 paired CRC and the adjacent normal tissues were detected by using SYBR Green real-time PCR method. Decreased expression of EphB6 protein was found in CRC as compared with adenoma and normal tissues (χ2 = 10.146, P = 0.001 and χ2 = 45.333, P < 0.001, respectively). Low EphB6 mRNA expression was detected in 83.8 % of cancers with negative or low EphB6 protein expression. The loss of EphB6 protein in CRC was positively associated with poorly differentiation (P < 0.001), lymph node metastasis (P = 0.006), Dukes stage (P = 0.002) and depth of invasion (P = 0.016). The patients with lymph node metastasis had a worse prognosis independently of gender, age, tumor site, stage and differentiation (RR = 0.404, CI 0.267–0.213, P < 0.001). Low levels of EphB6 protein expression are associated with a shorter mean duration of survival in colorectal cancer. Our results demonstrated that EphB6 may represent a novel, useful tissue biomarker for the prediction of survival rate in CRC.
Similar content being viewed by others
References
Ashktorab H, Schaffer AA, Daremipouran M, Smoot DT, Lee E, Brim H (2010) Distinct genetic alterations in colorectal cancer. PLoS One 5:e8879. doi:10.1371/journal.pone.0008879
Balakrishnan A et al (2007) Novel somatic and germline mutations in cancer candidate genes in glioblastoma, melanoma, and pancreatic carcinoma. Cancer Res 67:3545–3550. doi:10.1158/0008-5472.can-07-0065
Batlle E et al (2005) EphB receptor activity suppresses colorectal cancer progression. Nature 435:1126–1130. doi:10.1038/nature03626
Brantley-Sieders DM (2012) Clinical relevance of Ephs and ephrins in cancer: lessons from breast, colorectal, and lung cancer profiling. Semin Cell Dev Biol 23:102–108. doi:10.1016/j.semcdb.2011.10.014
Compton CC, Greene FL (2004) The staging of colorectal cancer: 2004 and beyond. CA Cancer J Clin 54:295–308
Coulthard MG et al (2002) The role of the Eph-ephrin signalling system in the regulation of developmental patterning. Int J Dev Biol 46:375–384
Ding L et al (2008) Somatic mutations affect key pathways in lung adenocarcinoma. Nature 455:1069–1075. doi:10.1038/nature07423
Elias D et al (2014) Prognostic similarities and differences in optimally resected liver metastases and peritoneal metastases from colorectal cancers. Ann Surg. doi:10.1097/SLA.0000000000000582
Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM (2010) Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 127:2893–2917. doi:10.1002/ijc.25516
Fox BP, Kandpal RP (2006) Transcriptional silencing of EphB6 receptor tyrosine kinase in invasive breast carcinoma cells and detection of methylated promoter by methylation specific PCR. Biochem Biophys Res Commun 340:268–276. doi:10.1016/j.bbrc.2005.11.174
Greenman C et al (2007) Patterns of somatic mutation in human cancer genomes. Nature 446:153–158. doi:10.1038/nature05610
Guo DL et al (2006) Reduced expression of EphB2 that parallels invasion and metastasis in colorectal tumours. Carcinogenesis 27:454–464. doi:10.1093/carcin/bgi259
Hafner C, Bataille F, Meyer S, Becker B, Roesch A, Landthaler M, Vogt T (2003) Loss of EphB6 expression in metastatic melanoma. Int J Oncol 23:1553–1559
Hafner C et al (2004) Differential gene expression of Eph receptors and ephrins in benign human tissues and cancers. Clin Chem 50:490–499. doi:10.1373/clinchem.2003.026849
Herath NI, Boyd AW (2010) The role of Eph receptors and ephrin ligands in colorectal cancer. Int J Cancer 126:2003–2011. doi:10.1002/ijc.25147
Herath NI, Doecke J, Spanevello MD, Leggett BA, Boyd AW (2009) Epigenetic silencing of EphA1 expression in colorectal cancer is correlated with poor survival. Br J Cancer 100:1095–1102. doi:10.1038/sj.bjc.6604970
Himanen JP (2012) Ectodomain structures of Eph receptors. Semin Cell Dev Biol 23:35–42. doi:10.1016/j.semcdb.2011.10.025
Kataoka H et al (2004) Correlation of EPHA2 overexpression with high microvessel count in human primary colorectal cancer. Cancer Sci 95:136–141
Kosinski C et al (2007) Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors. Proc Natl Acad Sci USA 104:15418–15423. doi:10.1073/pnas.0707210104
Kullander K, Klein R (2002) Mechanisms and functions of Eph and ephrin signalling. Nat Rev Mol Cell Biol 3:475–486. doi:10.1038/nrm856
Lisabeth EM, Falivelli G, Pasquale EB (2013) Eph receptor signaling and ephrins. Cold Spring Harb Perspect Biol 5. doi:10.1101/cshperspect.a009159
Menzel P, Valencia F, Godement P, Dodelet VC, Pasquale EB (2001) Ephrin-A6, a new ligand for EphA receptors in the developing visual system. Dev Biol 230:74–88. doi:10.1006/dbio.2000.0109
Merlos-Suarez A, Batlle E (2008) Eph-ephrin signalling in adult tissues and cancer. Curr Opin Cell Biol 20:194–200. doi:10.1016/j.ceb.2008.01.011
Migheli F, Migliore L (2012) Epigenetics of colorectal cancer. Clin Genet 81:312–318. doi:10.1111/j.1399-0004.2011.01829.x
Nakamura PA, Cramer KS (2013) EphB2 signaling regulates lesion-induced axon sprouting but not critical period length in the postnatal auditory brainstem. Neural Dev 8:2. doi:10.1186/1749-8104-8-2
Nikolov DB, Xu K, Himanen JP (2013) Eph/ephrin recognition and the role of Eph/ephrin clusters in signaling initiation. Biochim Biophys Acta 1834:2160–2165. doi:10.1016/j.bbapap.2013.04.020
Saika K, Sobue T (2013) Cancer statistics in the world. Gan To Kagaku Ryoho 40:2475–2480
Senior PV, Zhang BX, Chan ST (2010) Loss of cell-surface receptor EphB2 is important for the growth, migration, and invasiveness of a colon cancer cell line. Int J Colorectal Dis 25:687–694. doi:10.1007/s00384-010-0916-7
Sheng Z et al (2008) EphB1 is underexpressed in poorly differentiated colorectal cancers. Pathobiology 75:274–280. doi:10.1159/000151707
Walther A, Johnstone E, Swanton C, Midgley R, Tomlinson I, Kerr D (2009) Genetic prognostic and predictive markers in colorectal cancer. Nat Rev Cancer 9:489–499. doi:10.1038/nrc2645
Wang J et al (2005) Downregulation of EphA7 by hypermethylation in colorectal cancer. Oncogene 24:5637–5647. doi:10.1038/sj.onc.1208720
Xi HQ, Wu XS, Wei B, Chen L (2012) Eph receptors and ephrins as targets for cancer therapy. J Cell Mol Med 16:2894–2909. doi:10.1111/j.1582-4934.2012.01612.x
Yu J et al (2010) The EPHB6 receptor tyrosine kinase is a metastasis suppressor that is frequently silenced by promoter DNA hypermethylation in non-small cell lung cancer. Clin Cancer Res 16:2275–2283. doi:10.1158/1078-0432.ccr-09-2000
Acknowledgments
This work was supported in part by the National Natural Science Foundation of China (81371611, 81171391, 81372743) and the National Basic Research Priorities Program 973 Project (2014CB744504) from the Ministry of Science and Technology of China. We thank Prof. Fred Biddle for his help in modification of the manuscript.
Conflict of interest
The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Peng, L., Tu, P., Wang, X. et al. Loss of EphB6 protein expression in human colorectal cancer correlates with poor prognosis. J Mol Hist 45, 555–563 (2014). https://doi.org/10.1007/s10735-014-9577-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10735-014-9577-0