Abstract
Epithelial cell adhesion molecule (EpCAM) expression is elevated in breast cancer tissue, and correlates with the cancer metastasis and cell adhesion. Although EpCAM glycosylation is supposed to be associated with its function, the contribution of N-glycosylation to its function remains unclear. Here we analyzed cell adhesion ability of EpCAM in breast cancer cells. The results showed that EpCAM expression was associated with cell adhesion and N-glycosylation mutation of EpCAM decreased adhesion capacity. N-glycosylation mutation of EpCAM was correlated with lower levels of integrin β1 and fibronectin. We also found that effect of N-glycosylation of EpCAM on cell adhesion was regulated via FAK/Akt/Gsk-3β/β-catenin signaling pathway, which further adjusted MMP2/9 expression and activities. Our studies identified the characteristics and function of EpCAM glycosylation sites on breast cancer cell adhesion. These data could potentially clarify molecular regulation of EpCAM by N-glycosylation and intensify our understanding of the utility of glycosylated EpCAM as a target for breast cancer therapy.
Acknowledgments
This work was supported by grants from the Major State basic Research Development Program of China (2012CB822103), the National Natural Science Foundation of China (No.: 30800195, 31270866) and the National Natural Science Foundation of Liaoning Province (NO. 201602242).
References
Blandin, A. F., Renner, G., Lehmann, M., Lelong-Rebel, I., Martin, S., and Dontenwill, M. (2015). β1 Integrins as therapeutic targets to disrupt hallmarks of cancer. Front Pharmacol. 6, 279.10.3389/fphar.2015.00279Search in Google Scholar PubMed PubMed Central
Brunton, V. G. and Frame, M. C. (2008). Src and focal adhesion kinase as therapeutic targets in cancer. Curr. Opin. Pharmacol. 8, 427–432.10.1016/j.coph.2008.06.012Search in Google Scholar PubMed
Drake, R. R. (2015). Glycosylation and cancer: moving glycomics to the forefront. Adv. Cancer Res. 126, 1–10.10.1016/bs.acr.2014.12.002Search in Google Scholar PubMed
Gaiser, M. R., Lammermann, T., Feng, X., Igyarto, B. Z., Kaplan, D. H., Tessarollo, L., Germain, R. N., and Udey, M. C. (2012). Cancer-associated epithelial cell adhesion molecule (EpCAM; CD326) enables epidermal Langerhans cell motility and migration in vivo. Proc. Natl. Acad. Sci. USA. 109, E889–E897.10.1073/pnas.1117674109Search in Google Scholar PubMed PubMed Central
Gao, J., Liu, X., Yang, F., Liu, T., Yan, Q., and Yang, X. (2015). By inhibiting Ras/Raf/ERK and MMP-9, knockdown of EpCAM inhibits breast cancer cell growth and metastasis. Oncotarget 6, 27187–27198.10.18632/oncotarget.4551Search in Google Scholar PubMed PubMed Central
Gilboa-Geffen, A., Hamar, P., Le, M. T., Wheeler, L. A., Trifonova, R., Petrocca, F., Wittrup, A., and Lieberman, J. (2015). Gene knockdown by EpCAM aptamer-siRNA chimeras suppresses epithelial breast cancers and their tumor-initiating cells. Mol. Cancer. Ther. 14, 2279–2291.10.1158/1538-7445.AM2015-LB-299Search in Google Scholar
Gu, J. and Taniguchi, N. (2008). Potential of N-glycan in cell adhesion and migration as either a positive or negative regulator. Cell Adh. Migr. 2, 243–245.10.4161/cam.2.4.6748Search in Google Scholar PubMed PubMed Central
Guo, H. B., Johnson, H., Randolph, M., and Pierce, M. (2009). Regulation of homotypic cell-cell adhesion by branched N-glycosylation of N-cadherin extracellular EC2 and EC3 domains. J. Biol. Chem. 284, 34986–34997.10.1074/jbc.M109.060806Search in Google Scholar PubMed PubMed Central
Hauselmann, I. and Borsig, L. (2014). Altered tumor-cell glycosylation promotes metastasis. Front Oncol. 4, 28.10.3389/fonc.2014.00028Search in Google Scholar PubMed PubMed Central
Hiraga, T., Ito, S., and Nakamura, H. (2016). EpCAM expression in breast cancer cells is associated with enhanced bone metastasis formation. Int. J. Cancer 138, 1698–1708.10.1002/ijc.29921Search in Google Scholar PubMed
Isaji, T., Sato, Y., Fukuda, T., and Gu, J. (2009). N-glycosylation of the I-like domain of beta1 integrin is essential for β1 integrin expression and biological function: identification of the minimal N-glycosylation requirement for α5β1. J. Biol. Chem. 284, 12207–12216.10.1074/jbc.M807920200Search in Google Scholar
Isaji, T., Kariya, Y., Xu, Q., Fukuda, T., Taniguchi, N., and Gu, J. (2010). Functional roles of the bisecting GlcNAc in integrin-mediated cell adhesion. Methods Enzymol. 480, 445–459.10.1016/S0076-6879(10)80019-9Search in Google Scholar
Iwamoto, D. V. and Calderwood, D. A. (2015). Regulation of integrin-mediated adhesions. Curr. Opin. Cell Biol. 36, 41–47.10.1016/j.ceb.2015.06.009Search in Google Scholar PubMed PubMed Central
Kolbl, A. C., Andergassen, U., and Jeschke, U. (2015). The role of glycosylation in breast cancer metastasis and cancer control. Front Oncol. 5, 219.10.3389/fonc.2015.00219Search in Google Scholar PubMed PubMed Central
Lange, T., Samatov, T. R., Tonevitsky, A. G., and Schumacher, U. (2014). Importance of altered glycoprotein-bound N- and O-glycans for epithelial-to-mesenchymal transition and adhesion of cancer cells. Carbohydr. Res. 389, 39–45.10.1016/j.carres.2014.01.010Search in Google Scholar PubMed
Lannoo, N. and Van Damme, E. J. (2015). Review/N-glycans: the making of a varied toolbox. Plant. Sci. 239, 67–83.10.1016/j.plantsci.2015.06.023Search in Google Scholar PubMed
Li, W., Liu, Z., Zhao, C., and Zhai, L. (2015). Binding of MMP-9-degraded fibronectin to beta6 integrin promotes invasion via the FAK-Src-related Erk1/2 and PI3K/Akt/Smad-1/5/8 pathways in breast cancer. Oncol. Rep. 34, 1345–1352.10.3892/or.2015.4103Search in Google Scholar PubMed
Martowicz, A., Seeber, A., and Untergasser, G. (2016). The role of EpCAM in physiology and pathology of the epithelium. Histol. Histopathol. 31, 349–355.Search in Google Scholar
Mitra, S. K. and Schlaepfer, D. D. (2006). Integrin-regulated FAK-Src signaling in normal and cancer cells. Curr. Opin. Cell Biol. 18, 516–523.10.1016/j.ceb.2006.08.011Search in Google Scholar PubMed
Munz, M., Fellinger, K., Hofmann, T., Schmitt, B., and Gires, O. (2008). Glycosylation is crucial for stability of tumour and cancer stem cell antigen EpCAM. Front Biosci. 13, 5195–5201.10.2741/3075Search in Google Scholar PubMed
Nam, S. H., Kang, M., Ryu, J., Kim, H. J., Kim, D., Kim, D. G., Kwon, N. H., Kim, S., and Lee, J. W. (2016). Suppression of lysyl-tRNA synthetase, KRS, causes incomplete epithelial-mesenchymal transition and ineffective cellextracellular matrix adhesion for migration. Int. J. Oncol. 48, 1553–1560.10.3892/ijo.2016.3381Search in Google Scholar PubMed
Ohashi, R., Kawahara, K., Fujii, T., Takei, H., and Naito, Z. (2016). Higher expression of EpCAM is associated with poor clinical and pathological responses in breast cancer patients undergoing neoadjuvant chemotherapy. Pathol. Int. 66, 210–217.10.1111/pin.12404Search in Google Scholar
Pauli, C., Munz, M., Kieu, C., Mack, B., Breinl, P., Wollenberg, B., Lang, S., Zeidler, R., and Gires, O. (2003). Tumor-specific glycosylation of the carcinoma-associated epithelial cell adhesion molecule EpCAM in head and neck carcinomas. Cancer Lett. 193, 25–32.10.1016/S0304-3835(03)00003-XSearch in Google Scholar
Qin, J., Wang, L., Zheng, L., Zhou, X., Zhang, Y., Yang, T., and Zhou, Y. (2016). Concentrated growth factor promotes Schwann cell migration partly through the integrin beta1-mediated activation of the focal adhesion kinase pathway. Int. J. Mol. Med. 37, 1363–1370.10.3892/ijmm.2016.2520Search in Google Scholar PubMed
Schneck, H., Gierke, B., Uppenkamp, F., Behrens, B., Niederacher, D., Stoecklein, N. H., Templin, M. F., Pawlak, M., Fehm, T., Neubauer, H., et al. (2016). Correction: EpCAM-independent enrichment of circulating tumor cells in metastatic breast cancer. PLoS One 11, e0149315.10.1371/journal.pone.0149315Search in Google Scholar PubMed PubMed Central
Schnell, U., Cirulli, V., and Giepmans, B. N. (2013). EpCAM: structure and function in health and disease. Biochim. Biophys. Acta. 1828, 1989–2001.10.1016/j.bbamem.2013.04.018Search in Google Scholar PubMed
Stowell, S. R., Ju, T., and Cummings, R. D. (2015). Protein glycosylation in cancer. Annu. Rev. Pathol. 10, 473–510.10.1146/annurev-pathol-012414-040438Search in Google Scholar PubMed PubMed Central
Strasser, R. (2016). Plant protein glycosylation. Glycobiology 26, 926–939.10.1093/glycob/cww023Search in Google Scholar PubMed PubMed Central
Sulzmaier, F. J., Jean, C., and Schlaepfer, D. D. (2014). FAK in cancer: mechanistic findings and clinical applications. Nat. Rev. Cancer 14, 598–610.10.1038/nrc3792Search in Google Scholar PubMed PubMed Central
Taniguchi, N. and Kizuka, Y. (2015). Glycans and cancer: role of N-glycans in cancer biomarker, progression and metastasis, and therapeutics. Adv. Cancer Res. 126, 11–51.10.1016/bs.acr.2014.11.001Search in Google Scholar PubMed
Wang, S., Hu, J., Yao, Y., Shi, M., Yue, L., Han, F., Zhang, H., He, J., Liu, S., and Li, Y. (2013). MARVELD1 regulates integrin beta1-mediated cell adhesion and actin organization via inhibiting its pre-mRNA processing. Int. J. Biochem. Cell Biol. 45, 2679–2687.10.1016/j.biocel.2013.09.006Search in Google Scholar PubMed
Wangpu, X., Lu, J., Xi, R., Yue, F., Sahni, S., Park, K. C., Menezes, S., Huang, M. L., Zheng, M., Kovacevic, Z., et al. (2016). Targeting the metastasis suppressor, N-Myc downstream regulated Gene-1, with novel di-2- pyridylketone thiosemicarbazones: suppression of tumor cell migration and cell-collagen adhesion by inhibiting focal adhesion kinase/paxillin signaling. Mol. Pharmacol. 89, 521–540.10.1124/mol.115.103044Search in Google Scholar PubMed PubMed Central
Zhang, J. and Hochwald, S. N. (2014). The role of FAK in tumor metabolism and therapy. Pharmacol. Ther. 142, 154–163.10.1016/j.pharmthera.2013.12.003Search in Google Scholar PubMed PubMed Central
Zhang, Z. Y., Lu, Y. X., Zhang, Z. Y., Chang, Y. Y., Zheng, L., Yuan, L., Zhang, F., Hu, Y. H., Zhang, W. J., and Li, X. N. (2016). Loss of TINCR expression promotes proliferation, metastasis through activating EpCAM cleavage in colorectal cancer. Oncotarget 7, 22639–22649.10.18632/oncotarget.8141Search in Google Scholar PubMed PubMed Central
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