Ephrin‑B2 inhibits cell proliferation and motility in vitro and predicts longer metastasis‑free survival in breast cancer

Magic,Zeljana; Magic,Zeljana; Magic,Zeljana; Magic,Zeljana; Sandström,Josefine; Sandström,Josefine; Sandström,Josefine; Sandström,Josefine; Perez‑Tenorio,Gizeh; Perez‑Tenorio,Gizeh; Perez‑Tenorio,Gizeh; Perez‑Tenorio,Gizeh

doi:10.3892/ijo.2019.4892

Ephrin‑B2 inhibits cell proliferation and motility in vitro and predicts longer metastasis‑free survival in breast cancer

Authors:
- Zeljana Magic
- Josefine Sandström
- Gizeh Perez‑Tenorio
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Affiliations: Department of Clinical and Experimental Medicine and Department of Oncology, Linköping University, SE‑58185 Linköping, Sweden
Published online on: October 4, 2019 https://doi.org/10.3892/ijo.2019.4892
Pages: 1275-1286
Copyright: © Magic et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The tyrosine kinase receptor EphB4 and its ligand ephrin‑B2 interact through cell‑to‑cell contacts. Upon interaction, EphB4 transmits bidirectional signals. A forward signal inside EphB4‑expressing cells is believed to suppress tumor growth, while inside the ephrin‑expressing cells, an oncogenic reverse signal arises. In breast cancer cells with a high EphB4 receptor expression the forward signal is low, in part due to the low expression of the ligand ephrin‑B2. Therefore, we hypothesized that by re‑introducing the ligand in EphB4‑positive cells, tumor suppression could be induced by the stimulation of the forward signal. This question was addressed in vitro by the stable lentiviral infection of breast cancer cells with either wild‑type EFNB2 or with a mutant EFNB2‑5F, unable to transmit reverse signaling. Furthermore, we investigated ephrin‑B and EphB4 protein expression in 216 paraffin‑embedded tumors using immunohistochemistry. The in vitro results indicated that ephrin‑B2 expression was associated with a lower cell proliferation, migration and motility compared with the control cells. These effects were more pronounced when the cells lacked the ability to transmit the reverse signal (B2‑5F). In clinical material, ephrin‑B protein expression was associated with a positive estrogen receptor (ER) status, a low HER‑2 expression and was negatively associated with Nottingham histologic grade (NHG) III. Ephrin‑B expression indicated a good prognosis, whereas EphB4 expression was associated with a shorter metastasis‑free survival in univariate and multivariate analysis. Furthermore, the prognostic value of EFNB2 and EPHB4 was confirmed at the gene expression level in public datasets. Thus, on the whole, the findings of this study suggest that ephrin‑B2 expression is associated with less proliferation and lower motility of breast cancer cells and with a longer patient survival in breast cancer.

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1	Pasquale EB: Eph receptors and ephrins in cancer: Bidirectional signalling and beyond. Nat Rev Cancer. 10:165–180. 2010. View Article : Google Scholar : PubMed/NCBI
2	Vaught D, Brantley-Sieders DM and Chen J: Eph receptors in breast cancer: Roles in tumor promotion and tumor suppression. Breast Cancer Res. 10:2172008. View Article : Google Scholar
3	Hirai H, Maru Y, Hagiwara K, Nishida J and Takaku F: A novel putative tyrosine kinase receptor encoded by the eph gene. Science. 238:1717–1720. 1987. View Article : Google Scholar : PubMed/NCBI
4	Dodelet VC and Pasquale EB: Eph receptors and ephrin ligands: Embryogenesis to tumorigenesis. Oncogene. 19:5614–5619. 2000. View Article : Google Scholar : PubMed/NCBI
5	Zhuang G, Brantley-Sieders DM, Vaught D, Yu J, Xie L, Wells S, Jackson D, Muraoka-Cook R, Arteaga C and Chen J: Elevation of receptor tyrosine kinase EphA2 mediates resistance to trastu-zumab therapy. Cancer Res. 70:299–308. 2010. View Article : Google Scholar
6	Brantley-Sieders DM, Jiang A, Sarma K, Badu-Nkansah A, Walter DL, Shyr Y and Chen J: Eph/ephrin profiling in human breast cancer reveals significant associations between expression level and clinical outcome. PLoS One. 6:e244262011. View Article : Google Scholar : PubMed/NCBI
7	Lu M, Miller KD, Gokmen-Polar Y, Jeng MH and Kinch MS: EphA2 overexpression decreases estrogen dependence and tamoxifen sensitivity. Cancer Res. 63:3425–3429. 2003.PubMed/NCBI
8	Wu Q, Suo Z, Risberg B, Karlsson MG, Villman K and Nesland JM: Expression of Ephb2 and Ephb4 in breast carcinoma. Pathol Oncol Res. 10:26–33. 2004. View Article : Google Scholar : PubMed/NCBI
9	Zelinski DP, Zantek ND, Stewart JC, Irizarry AR and Kinch MS: EphA2 overexpression causes tumorigenesis of mammary epithelial cells. Cancer Res. 61:2301–2306. 2001.PubMed/NCBI
10	Brantley-Sieders DM: Clinical relevance of Ephs and ephrins in cancer: Lessons from breast, colorectal, and lung cancer profiling. Semin Cell Dev Biol. 23:102–108. 2012. View Article : Google Scholar :
11	Kumar SR, Singh J, Xia G, Krasnoperov V, Hassanieh L, Ley EJ, Scehnet J, Kumar NG, Hawes D, Press MF, et al: Receptor tyrosine kinase EphB4 is a survival factor in breast cancer. Am J Pathol. 169:279–293. 2006. View Article : Google Scholar : PubMed/NCBI
12	Noren NK, Foos G, Hauser CA and Pasquale EB: The EphB4 receptor suppresses breast cancer cell tumorigenicity through an Abl-Crk pathway. Nat Cell Biol. 8:815–825. 2006. View Article : Google Scholar : PubMed/NCBI
13	Berclaz G, Flütsch B, Altermatt HJ, Rohrbach V, Djonov V, Ziemiecki A, Dreher E and Andres AC: Loss of EphB4 receptor tyrosine kinase protein expression during carcinogenesis of the human breast. Oncol Rep. 9:985–989. 2002.PubMed/NCBI
14	Nikolova Z, Djonov V, Zuercher G, Andres AC and Ziemiecki A: Cell-type specific and estrogen dependent expression of the receptor tyrosine kinase EphB4 and its ligand ephrin-B2 during mammary gland morphogenesis. J Cell Sci. 111:2741–2751. 1998.PubMed/NCBI
15	Mäkinen T, Adams RH, Bailey J, Lu Q, Ziemiecki A, Alitalo K, Klein R and Wilkinson GA: PDZ interaction site in ephrinB2 is required for the remodeling of lymphatic vasculature. Genes Dev. 19:397–410. 2005. View Article : Google Scholar : PubMed/NCBI
16	Salvucci O, Maric D, Economopoulou M, Sakakibara S, Merlin S, Follenzi A and Tosato G: EphrinB reverse signaling contributes to endothelial and mural cell assembly into vascular structures. Blood. 114:1707–1716. 2009. View Article : Google Scholar : PubMed/NCBI
17	Follenzi A and Naldini L: Generation of HIV-1 derived lentiviral vectors. Methods Enzymol. 346:454–465. 2002. View Article : Google Scholar : PubMed/NCBI
18	Falivelli G, Lisabeth EM, Rubio de la Torre E, Perez-Tenorio G, Tosato G, Salvucci O and Pasquale EB: Attenuation of eph receptor kinase activation in cancer cells by coexpressed ephrin ligands. PLoS One. 8:e814452013. View Article : Google Scholar : PubMed/NCBI
19	Rutqvist LE and Johansson H: Long-term follow-up of the Stockholm randomized trials of postoperative radiation therapy versus adjuvant chemotherapy among 'high risk' pre- and postmenopausal breast cancer patients. Acta Oncol. 45:517–527. 2006. View Article : Google Scholar : PubMed/NCBI
20	Veenstra C, Pérez-Tenorio G, Stelling A, Karlsson E, Mirwani SM, Nordensköljd B and Fornander T: Met and its ligand HGF are associated with clinical outcome in breast cancer. Oncotarget. 7:37145–37159. 2016. View Article : Google Scholar : PubMed/NCBI
21	Husa AM, Magić Ž, Larsson M, Fornander T and Pérez-Tenorio G: EPH/ephrin profile and EPHB2 expression predicts patient survival in breast cancer. Oncotarget. 7:21362–21380. 2016. View Article : Google Scholar : PubMed/NCBI
22	Pawitan Y, Bjöhle J, Amler L, Borg AL, Egyhazi S, Hall P, Han X, Holmberg L, Huang F, Klaar S, et al: Gene expression profiling spares early breast cancer patients from adjuvant therapy: Derived and validated in two population-based cohorts. Breast Cancer Res. 7:R953–R964. 2005. View Article : Google Scholar : PubMed/NCBI
23	van de Vijver MJ, He YD, van't Veer LJ, Dai H, Hart AA, Voskuil DW, Schreiber GJ, Peterse JL, Roberts C, Marton MJ, et al: A gene-expression signature as a predictor of survival in breast cancer. N Engl J Med. 347:1999–2009. 2002. View Article : Google Scholar : PubMed/NCBI
24	Ruan SQ, Wang SW, Wang ZH and Zhang SZ: Regulation of HRG-β1-induced proliferation, migration and invasion of MCF-7 cells by upregulation of GPR30 expression. Mol Med Rep. 6:131–138. 2012.PubMed/NCBI
25	Mezi S, Todi L, Orsi E, Angeloni A and Mancini P: Involvement of the Src-cortactin pathway in migration induced by IGF-1 and EGF in human breast cancer cells. Int J Oncol. 41:2128–2138. 2012. View Article : Google Scholar : PubMed/NCBI
26	Rutkowski R, Mertens-Walker I, Lisle JE, Herington AC and Stephenson SA: Evidence for a dual function of EphB4 as tumor promoter and suppressor regulated by the absence or presence of the ephrin-B2 ligand. Int J Cancer. 131:E614–E624. 2012. View Article : Google Scholar
27	Alam SM, Fujimoto J, Jahan I, Sato E and Tamaya T: Coexpression of EphB4 and ephrinB2 in tumor advancement of uterine cervical cancers. Gynecol Oncol. 114:84–88. 2009. View Article : Google Scholar : PubMed/NCBI
28	Liu W, Ahmad SA, Jung YD, Reinmuth N, Fan F, Bucana CD and Ellis LM: Coexpression of ephrin-Bs and their receptors in colon carcinoma. Cancer. 94:934–939. 2002. View Article : Google Scholar : PubMed/NCBI
29	Tang XX, Brodeur GM, Campling BG and Ikegaki N: Coexpression of transcripts encoding EPHB receptor protein tyrosine kinases and their ephrin-B ligands in human small cell lung carcinoma. Clin Cancer Res. 5:455–460. 1999.PubMed/NCBI
30	Noberini R, Rubio de la Torre E and Pasquale EB: Profiling Eph receptor expression in cells and tissues: A targeted mass spectrometry approach. Cell Adh Migr. 6:102–112. 2012. View Article : Google Scholar : PubMed/NCBI
31	Foo SS, Turner CJ, Adams S, Compagni A, Aubyn D, Kogata N, Lindblom P, Shani M, Zicha D and Adams RH: Ephrin-B2 controls cell motility and adhesion during blood-vessel-wall assembly. Cell. 124:161–173. 2006. View Article : Google Scholar : PubMed/NCBI
32	Bochenek ML, Dickinson S, Astin JW, Adams RH and Nobes CD: Ephrin-B2 regulates endothelial cell morphology and motility independently of Eph-receptor binding. J Cell Sci. 123:1235–1246. 2010. View Article : Google Scholar : PubMed/NCBI
33	Guijarro-Muñoz I, Sánchez A, Martínez-Martínez E, García JM, Salas C, Provencio M, Alvarez-Vallina L and Sanz L: Gene expression profiling identifies EPHB4 as a potential predictive biomarker in colorectal cancer patients treated with bevacizumab. Med Oncol. 30:5722013. View Article : Google Scholar : PubMed/NCBI
34	Tu Y, He S, Fu J, Li G, Xu R, Lu H and Deng J: Expression of EphrinB2 and EphB4 in glioma tissues correlated to the progression of glioma and the prognosis of glioblastoma patients. Clin Transl Oncol. 14:214–220. 2012. View Article : Google Scholar : PubMed/NCBI
35	Cancer Genome Atlas Network: Comprehensive molecular portraits of human breast tumours. Nature. 490:61–70. 2012. View Article : Google Scholar : PubMed/NCBI
36	Pereira B, Chin SF, Rueda OM, Vollan HK, Provenzano E, Bardwell HA, Pugh M, Jones L, Russell R, Sammut SJ, et al: The somatic mutation profiles of 2,433 breast cancers refines their genomic and transcriptomic landscapes. Nat Commun. 7:114792016. View Article : Google Scholar : PubMed/NCBI