Silencing of EPHB2 promotes the epithelial‑mesenchymal transition of skin squamous cell carcinoma‑derived A431 cells

Inagaki,Yoshinori; Tokunaga,Tomohiko; Yanai,Mitsuru; Wu,Dan; Huang,Jiyi; Nagase,Hiroki; Fukuda,Noboru; Ozaki,Toshinori; Soma,Masayoshi; Fujiwara,Kyoko

doi:10.3892/ol.2019.10019

Silencing of EPHB2 promotes the epithelial‑mesenchymal transition of skin squamous cell carcinoma‑derived A431 cells

Authors:
- Yoshinori Inagaki
- Tomohiko Tokunaga
- Mitsuru Yanai
- Dan Wu
- Jiyi Huang
- Hiroki Nagase
- Noboru Fukuda
- Toshinori Ozaki
- Masayoshi Soma
- Kyoko Fujiwara
View Affiliations

Affiliations: Division of General Medicine, Department of Medicine, Nihon University School of Medicine, Itabashi, Tokyo 173‑8610, Japan, Department of Oncology, The 5th Hospital of Xiamen, Xiamen, Fujian 361101, P.R. China, Department of Nephropathy, The 5th Hospital of Xiamen, Xiamen, Fujian 361101, P.R. China, Laboratory of Cancer Genetics, Chiba Cancer Center Research Institute, Chiba 260‑8717, Japan, Division of Nephrology, Hypertension and Endocrinology, Department of Medicine, Nihon University School of Medicine, Itabashi, Tokyo 173‑8610, Japan, Department of DNA Damage Signaling, Research Center, The 5th Hospital of Xiamen, Xiamen, Fujian 361101, P.R. China
Published online on: February 6, 2019 https://doi.org/10.3892/ol.2019.10019
Pages: 3735-3742
Copyright: © Inagaki et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Erythropoietin‑producing hepatocellular (Eph) receptors and their ligand ephrins serve crucial roles in the interactions among epithelial cells. Eph receptor/ephrin signaling regulates cell functions, including proliferation, differentiation and migration, via these cell‑cell interactions. We reported previously that EPHB2, a member of the Eph receptor family, was highly expressed in chemically induced cutaneous squamous cell carcinoma (cSCC) tissues in mice. Although the higher expression level of EPHB2 has been observed in various human cancers, its roles in the development and progression of cancers are still unclear. In the present study, the functional implications of EPHB2 in the acquisition of malignant phenotypes of cSCC cells was investigated. Silencing of EPHB2 in the human cSCC cell line A431 induced epithelial‑mesenchymal transition (EMT)‑like morphological changes accompanied by a significant upregulation of epithelial‑mesenchymal transition‑associated genes such as zinc finger E‑box binding homeobox 1/2. In addition, silencing of EPHB2 suppressed anchorage‑independent cell growth under 3D culture conditions. Consistent with these observations, EPHB2 exhibited higher levels of expression in tumor spheres formed under 3D culture conditions than in cells cultured in adherent form, and the expression pattern of EMT markers indicated that EMT was suppressed in tumor spheres. The results of the present study indicated that EPHB2 serves a pivotal role in promoting the anchorage‑independent growth of A431 cells through the suppression of EMT.

View Figures

View References

1	Pasquale EB: Eph-ephrin bidirectional signaling in physiology and disease. Cell. 133:38–52. 2008. View Article : Google Scholar : PubMed/NCBI
2	Gale NW, Holland SJ, Valenzuela DM, Flenniken A, Pan L, Ryan TE, Henkemeyer M, Strebhardt K, Hirai H, Wilkinson DG, et al: Eph receptors and ligands comprise two major specificity subclasses and are reciprocally compartmentalized during embryogenesis. Neuron. 17:9–19. 1996. View Article : Google Scholar : PubMed/NCBI
3	Wijeratne DT, Rodger J, Wood FM and Fear MW: The role of Eph receptors and Ephrins in the skin. Int J Dermatol. 55:3–10. 2016. View Article : Google Scholar : PubMed/NCBI
4	Pasquale EB: Eph receptor signalling casts a wide net on cell behaviour. Nat Rev Mol Cell Biol. 6:462–475. 2005. View Article : Google Scholar : PubMed/NCBI
5	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
6	Gao Q, Liu W, Cai J, Li M, Gao Y, Lin W and Li Z: EphB2 promotes cervical cancer progression by inducing epithelial-mesenchymal transition. Hum Pathol. 45:372–381. 2014. View Article : Google Scholar : PubMed/NCBI
7	Kumar SR, Scehnet JS, Ley EJ, Singh J, Krasnoperov V, Liu R, Manchanda PK, Ladner RD, Hawes D, Weaver FA, et al: Preferential induction of EphB4 over EphB2 and its implication in colorectal cancer progression. Cancer Res. 69:3736–3745. 2009. View Article : Google Scholar : PubMed/NCBI
8	Kumar SR, Masood R, Spannuth WA, Singh J, Scehnet J, Kleiber G, Jennings N, Deavers M, Krasnoperov V, Dubeau L, et al: The receptor tyrosine kinase EphB4 is overexpressed in ovarian cancer, provides survival signals and predicts poor outcome. Br J Cancer. 96:1083–1091. 2007. View Article : Google Scholar : PubMed/NCBI
9	Hafner C, Becker B, Landthaler M and Vogt T: Expression profile of Eph receptors and ephrin ligands in human skin and downregulation of EphA1 in nonmelanoma skin cancer. Mod Pathol. 19:1369–1377. 2006. View Article : Google Scholar : PubMed/NCBI
10	Herath NI, Doecke J, Spanevello MD, Leggett BA and Boyd AW: Epigenetic silencing of EphA1 expression in colorectal cancer is correlated with poor survival. Br J Cancer. 100:1095–1102. 2009. View Article : Google Scholar : PubMed/NCBI
11	Batlle E, Bacani J, Begthel H, Jonkheer S, Gregorieff A, van de Born M, Malats N, Sancho E, Boon E, Pawson T, et al: EphB receptor activity suppresses colorectal cancer progression. Nature. 435:1126–1130. 2005. View Article : Google Scholar : PubMed/NCBI
12	Li JJ, Liu DP, Liu GT and Xie D: EphrinA5 acts as a tumor suppressor in glioma by negative regulation of epidermal growth factor receptor. Oncogene. 28:1759–1768. 2009. View Article : Google Scholar : PubMed/NCBI
13	Fujiwara K, Ghosh S, Liang P, Morien E, Soma M and Nagase H: Genome-wide screening of aberrant DNA methylation which associated with gene expression in mouse skin cancers. Mol Carcinog. 54:178–188. 2015. View Article : Google Scholar : PubMed/NCBI
14	Farshchian M, Nissinen L, Siljamäki E, Riihilä P, Toriseva M, Kivisaari A, Ala-Aho R, Kallajoki M, Veräjänkorva E, Honkanen HK, et al: EphB2 promotes progression of cutaneous squamous cell carcinoma. J Invest Dermatol. 135:1882–1892. 2015. View Article : Google Scholar : PubMed/NCBI
15	Li RX, Chen ZH and Chen ZK: The role of EPH receptors in cancer-related epithelial-mesenchymal transition. Chin J Cancer. 33:231–240. 2014. View Article : Google Scholar : PubMed/NCBI
16	Savagner P: The epithelial-mesenchymal transition (EMT) phenomenon. Ann Oncol. 21 (Suppl 7):vii89–vii92. 2010. View Article : Google Scholar : PubMed/NCBI
17	Yuan W and Chen Z, Wu S, Ge J, Chang S, Wang X, Chen J and Chen Z: Expression of EphA2 and E-cadherin in gastric cancer: Correlated with tumor progression and lymphogenous metastasis. Pathol Oncol Res. 15:473–478. 2009. View Article : Google Scholar : PubMed/NCBI
18	Saito T, Masuda N, Miyazaki T, Kanoh K, Suzuki H, Shimura T, Asao T and Kuwano H: Expression of EphA2 and E-cadherin in colorectal cancer: Correlation with cancer metastasis. Oncol Rep. 11:605–611. 2004.PubMed/NCBI
19	Schauer MC, Stoecklein NH, Theisen J, Kropil F, Baldus S, Hoelscher A, Feith M, Bolke E, Matuschek C, Budach W and Knoefel WT: The simultaneous expression of both ephrin B3 receptor and E-cadherin in Barrett's adenocarcinoma is associated with favorable clinical staging. Eur J Med Res. 17:102012. View Article : Google Scholar : PubMed/NCBI
20	Zhou P, Li B, Liu F, Zhang M, Wang Q, Liu Y, Yao Y and Li D: The epithelial to mesenchymal transition (EMT) and cancer stem cells: implication for treatment resistance in pancreatic cancer. Mol Cancer. 16:522017. View Article : Google Scholar : PubMed/NCBI
21	Izumiya M, Kabashima A, Higuchi H, Igarashi T, Sakai G, Iizuka H, Nakamura S, Adachi M, Hamamoto Y, Funakoshi S, et al: Chemoresistance is associated with cancer stem cell-like properties and epithelial-to-mesenchymal transition in pancreatic cancer cells. Anticancer Res. 32:3847–3853. 2012.PubMed/NCBI
22	Guadamillas MC, Cerezo A and Del Pozo MA: Overcoming anoikis-pathways to anchorage-independent growth in cancer. J Cell Sci. 124:3189–3197. 2011. View Article : Google Scholar : PubMed/NCBI
23	Malaguarnera R and Belfiore A: The emerging role of insulin and insulin-like growth factor signaling in cancer stem cells. Front Endocrinol (Lausanne). 5:102014. View Article : Google Scholar : PubMed/NCBI
24	Lam S, Wiercinska E, Teunisse AF, Lodder K, ten Dijke P and Jochemsen AG: Wild-type p53 inhibits pro-invasive properties of TGF-β3 in breast cancer, in part through regulation of EPHB2, a new TGF-β target gene. Breast Cancer Res Treat. 148:7–18. 2014. View Article : Google Scholar : PubMed/NCBI
25	Harlow E, Williamson NM, Ralston R, Helfman DM and Adams TE: Molecular cloning and in vitro expression of a cDNA clone for human cellular tumor antigen p53. Mol Cell Biol. 5:1601–1610. 1985. View Article : Google Scholar : PubMed/NCBI