表皮生長因子的活化是許多癌症造成癌轉移的主要原因，其中像是頭頸鱗狀癌細胞。一種叫做纖維連接蛋白的的多功能細胞外醣蛋白在細胞的黏附、生長、分化以遷移扮演很重要的角色。在我們實驗室先前的研究發現，剔除環氧合酶2 (cyclooxygenase-2)的表現會抑制掉表皮生長因子所誘導的纖維連接蛋白表現以及纖維連接蛋白所導致的腫瘤轉移。然而在頭頸癌細胞裡表皮生長因子誘導纖維連接蛋白這其中的機制以及癌轉移仍尚未瞭解。我們首先確認了表皮生長因子確實可以促進頭頸癌細胞裡纖維連接蛋白的mRNA和蛋白表現。下一步我們證實了表皮生長因子調控纖維連接蛋白的表現是透過穩定mRNA的穩定性而不是透過基因轉錄。而我們確定在表皮生長因子穩定纖維連接蛋白的mRNA的穩定性的機制是透纖維連接蛋白的五端非轉譯區所調控。接著我們發現異質核核醣核蛋白A2/B1的過度表現會增加纖維連接蛋白的蛋白表現以及基質金屬蛋白酶2的mRNA表現。此外，剔除掉核仁素的基因表現會去抑制纖維連接蛋白的mRNA穩定性、mRNA以及蛋白表現。除此之外，我們也發現表皮生長因子會透過NF-κB和ERK的訊息傳遞路徑來調控核仁素進而影響纖維連接蛋白的表現。

The aim of this study is to characterize whether EGF can induce Fibronectin expression and investigate the function role about fibronectin in tumor metastasis. In the present study, we used real-time PCR and western blotting to clarify that EGF-induced the expression of fibronectin was in a time-dependent manner in FaDu, TU183 and HONE-1 cells. We found that EGF induces fibronectin expression through stabilizing fibronectin mRNA by actinomycin D treatment. We found that EGF regulated fibronectin mRNA stability through fibronectin 5’ untranslated region. The RNA-binding protein, hnRNAPA2B1 and nucleolin regulated fibronectin mRNA and protein expression and fibronectin mRNA stability. In addition, we also found that EGF regulated fibronectin through NF-κB and ERK signaling pathways.

Abdelmohsen, K., & Gorospe, M. (2012). RNA-binding protein nucleolin in disease. RNA Biol, 9(6), 799-808. doi: 10.4161/rna.19718
Akiyama SK, Olden K, Yamada KM. (1995). Fibronectin and integrins in invasion and metastasis. Cancer Metastasis Rev., 14, 173-189.
Aota S., Nomizu M., Yamada K.M. (1994). The short amino acid sequence Pro-His-Ser-Arg-Asn in human fibronectin enhances cell-adhesive function. . J. Biol. Chem., 269, 24756-24761.
Cao, Y., Liu, X., Lu, W., Chen, Y., Wu, X., Li, M., . . . Liu, Y. (2015). Fibronectin promotes cell proliferation and invasion through mTOR signaling pathway activation in gallbladder cancer. Cancer Lett, 360(2), 141-150. doi: 10.1016/j.canlet.2015.01.041
Chang, W. C., Wu, S. L., Huang, W. C., Hsu, J. Y., Chan, S. H., Wang, J. M., . . . Chen, B. K. (2015). PTX3 gene activation in EGF-induced head and neck cancer cell metastasis. Oncotarget, 6(10), 7741-7757. doi: 10.18632/oncotarget.3482
Chen, I. H., Chang, J. T., Liao, C. T., Wang, H. M., Hsieh, L. L., & Cheng, A. J. (2003). Prognostic significance of EGFR and Her-2 in oral cavity cancer in betel quid prevalent area cancer prognosis. Br J Cancer, 89(4), 681-686. doi: 10.1038/sj.bjc.6601171
Deane F. Mosher, M.D. (1984). Physiology of fibronectin Annual Reviews Inc., 561-575.
Deng, J., Chen, S., Wang, F., Zhao, H., Xie, Z., Xu, Z., . . . Cheng, Y. (2016). Effects of hnRNP A2/B1 Knockdown on Inhibition of Glioblastoma Cell Invasion, Growth and Survival. Mol Neurobiol, 53(2), 1132-1144. doi: 10.1007/s12035-014-9080-3
Dunn, G. P., Old, L. J., & Schreiber, R. D. (2004). The immunobiology of cancer immunosurveillance and immunoediting. Immunity, 21(2), 137-148. doi: 10.1016/j.immuni.2004.07.017
Duvvuri, Umamaheswar, & Myers, Jeffrey N. (2009). Cancer of the Head and Neck Is the Sixth Most Common Cancer Worldwide. Curr Probl Surg 46(2), 114-117.
Garcia-Blanco, Alex M. WardJ. GunaratneMariano A. (2014). Identification of Dengue RNA Binding Proteins Using RNA Chromatography and Quantitative Mass Spectrometry. Dengue, 253-270.
Geuens, T., Bouhy, D., & Timmerman, V. (2016). The hnRNP family: insights into their role in health and disease. Hum Genet, 135(8), 851-867. doi: 10.1007/s00439-016-1683-5
Gillison ML, Koch WM, Capone RB, Spafford M, Westra WH, Wu L, Zahurak ML, Daniel RW, Viglione M, Symer DE, Shah KV, Sidransky D. (2000). Evidence for a causal association between human papillomavirus and a subset of head and neck cancers. J Natl Cancer Inst., 92(9), 709-720.
Goldstein., John Decker. Jerome C. (1982). Risk Factors in Head and Neck Cancer. N Engl J Med 306(19), 1151-1155.
Grandis, Rebecca G. PomerantzJennifer Rubin. (2003). The role of epidermal growth factor receptor in head and neck squamous cell carcinoma. Oncol Rep, 5, 140-146.
Hanahan, D., & Weinberg, R. A. (2011). Hallmarks of cancer: the next generation. Cell, 144(5), 646-674. doi: 10.1016/j.cell.2011.02.013
Hervé Ginisty, Hélène Sicard, Benoit Roger and Philippe Bouvet. (1999). Structure and functions of nucleolin. Journal of Cell Science, 112, 761-772.
Higashiyama, S., Abraham, J. A., Miller, J., Fiddes, J. C., & Klagsbrun, M. (1991). A heparin-binding growth factor secreted by macrophage-like cells that is related to EGF. Science, 251(4996), 936-939.
Hsu, J. Y., Chang, J. Y., Chang, K. Y., Chang, W. C., & Chen, B. K. (2017). Epidermal growth factor-induced pyruvate dehydrogenase kinase 1 expression enhances head and neck squamous cell carcinoma metastasis via up-regulation of fibronectin. FASEB J. doi: 10.1096/fj.201700156R
Hsu, J. Y., Chang, K. Y., Chen, S. H., Lee, C. T., Chang, S. T., Cheng, H. C., . . . Chen, B. K. (2015). Epidermal growth factor-induced cyclooxygenase-2 enhances head and neck squamous cell carcinoma metastasis through fibronectin up-regulation. Oncotarget, 6(3), 1723-1739. doi: 10.18632/oncotarget.2783
Hsu, T. I., Lin, S. C., Lu, P. S., Chang, W. C., Hung, C. Y., Yeh, Y. M., . . . Hung, J. J. (2015). MMP7-mediated cleavage of nucleolin at Asp255 induces MMP9 expression to promote tumor malignancy. Oncogene, 34(7), 826-837. doi: 10.1038/onc.2014.22
Hynes, N. E., & Lane, H. A. (2005). ERBB receptors and cancer: the complexity of targeted inhibitors. Nat Rev Cancer, 5(5), 341-354. doi: 10.1038/nrc1609
Kalyankrishna, S., & Grandis, J. R. (2006). Epidermal growth factor receptor biology in head and neck cancer. J Clin Oncol, 24(17), 2666-2672. doi: 10.1200/JCO.2005.04.8306
Kerrebijn, J. D., Balm, A. J., Knegt, P. P., Meeuwis, C. A., & Drexhage, H. A. (1994). Macrophage and dendritic cell infiltration in head and neck squamous-cell carcinoma; an immunohistochemical study. Cancer Immunol Immunother, 38(1), 31-37.
Kimizuka F., Ohdate Y., Kawase Y., Shimojo T., Taguchi Y., Hashino K., Goto S., Hashi H., Kato I., Sekiguchi K., Titani K. (1991). Role of Type I11 Homology Repeats in Cell Adhesive Function within the Cell-binding Domain of Fibronectin. J. Biol. Chem., 266, 3045-3051.
Kuhn, A. N., Diken, M., Kreiter, S., Selmi, A., Kowalska, J., Jemielity, J., . . . Sahin, U. (2010). Phosphorothioate cap analogs increase stability and translational efficiency of RNA vaccines in immature dendritic cells and induce superior immune responses in vivo. Gene Ther, 17(8), 961-971. doi: 10.1038/gt.2010.52
Lai, C. H., Tseng, J. T., Lee, Y. C., Chen, Y. J., Lee, J. C., Lin, B. W., . . . Hung, L. Y. (2010). Translational up-regulation of Aurora-A in EGFR-overexpressed cancer. J Cell Mol Med, 14(6B), 1520-1531. doi: 10.1111/j.1582-4934.2009.00919.x
Lee, P. T., Liao, P. C., Chang, W. C., & Tseng, J. T. (2007). Epidermal growth factor increases the interaction between nucleolin and heterogeneous nuclear ribonucleoprotein K/poly(C) binding protein 1 complex to regulate the gastrin mRNA turnover. Mol Biol Cell, 18(12), 5004-5013. doi: 10.1091/mbc.E07-04-0384
Liao, C. T., Chang, J. T., Wang, H. M., Ng, S. H., Hsueh, C., Lee, L. Y., . . . Yen, T. C. (2008). Analysis of risk factors of predictive local tumor control in oral cavity cancer. Ann Surg Oncol, 15(3), 915-922. doi: 10.1245/s10434-007-9761-5
Liao, C. T., Wang, H. M., Chang, J. T., Ng, S. H., Hsueh, C., Lee, L. Y., . . . Yen, T. C. (2007). Analysis of risk factors for distant metastases in squamous cell carcinoma of the oral cavity. Cancer, 110(7), 1501-1508. doi: 10.1002/cncr.22959
Lorch, J. H., Klessner, J., Park, J. K., Getsios, S., Wu, Y. L., Stack, M. S., & Green, K. J. (2004). Epidermal growth factor receptor inhibition promotes desmosome assembly and strengthens intercellular adhesion in squamous cell carcinoma cells. J Biol Chem, 279(35), 37191-37200. doi: 10.1074/jbc.M405123200
Lu, Z., Jiang, G., Blume-Jensen, P., & Hunter, T. (2001). Epidermal growth factor-induced tumor cell invasion and metastasis initiated by dephosphorylation and downregulation of focal adhesion kinase. Mol Cell Biol, 21(12), 4016-4031. doi: 10.1128/MCB.21.12.4016-4031.2001
Mao, L., Hong, W. K., & Papadimitrakopoulou, V. A. (2004). Focus on head and neck cancer. Cancer Cell, 5(4), 311-316.
Mao, Y., & Schwarzbauer, J. E. (2005). Fibronectin fibrillogenesis, a cell-mediated matrix assembly process. Matrix Biol, 24(6), 389-399. doi: 10.1016/j.matbio.2005.06.008
Maria Sundvall, Anna Karrila, Janne Nordberg, Reidar Grénman & Klaus Elenius. (2010). EGFR targeting drugs in the treatment of head and neck squamous cell carcinoma. J Expert Opinion on Emerging Drugs 15(2), 185-201.
Mimura, Y., Ihn, H., Jinnin, M., Asano, Y., Yamane, K., & Tamaki, K. (2004). Epidermal growth factor induces fibronectin expression in human dermal fibroblasts via protein kinase C delta signaling pathway. J Invest Dermatol, 122(6), 1390-1398. doi: 10.1111/j.0022-202X.2004.22618.x
Mosesson, M. (1970). The cold insoluble globulin of human plasma part 1 purification primary characterization and relationship to fibrinogen and other cold insoluble fraction components. Journal of Biological Chemistry, 245, 5728-5736.
Obara M., Kang M.S., Yamada K. (1988). Site-directed mutagenesis of the cell-binding domain of human fibronectinseparable, synergistic sites mediate adhesive function., 55, 649-657.
Orom, U. A., Nielsen, F. C., & Lund, A. H. (2008). MicroRNA-10a binds the 5'UTR of ribosomal protein mRNAs and enhances their translation. Mol Cell, 30(4), 460-471. doi: 10.1016/j.molcel.2008.05.001
Pankov, R. (2002). Fibronectin at a glance. Journal of Cell Science, 115(20), 3861-3863. doi: 10.1242/jcs.00059
Pierschbacher MD, Ruoslahti E. (1984). Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature, 309, 30-33.
PR, MORRISON. (1948). Preparation and properties of serum and plasma proteins; the separation of purified fibrinogen from fraction I of human plasma. J Am Chem Soc., 70, 3103-3108.
Reuter, C. W., Morgan, M. A., & Eckardt, A. (2007). Targeting EGF-receptor-signalling in squamous cell carcinomas of the head and neck. Br J Cancer, 96(3), 408-416. doi: 10.1038/sj.bjc.6603566
Richter, P., Umbreit, C., Franz, M., Berndt, A., Grimm, S., Uecker, A., . . . Berndt, A. (2011). EGF/TGFbeta1 co-stimulation of oral squamous cell carcinoma cells causes an epithelial-mesenchymal transition cell phenotype expressing laminin 332. J Oral Pathol Med, 40(1), 46-54. doi: 10.1111/j.1600-0714.2010.00936.x
RO, Hynes. (1973). Alteration of cell-surface proteins by viral transformation and by proteol. Proc Natl Acad Sci 70, 3170-3174.
Ruoslahti E, Vaheri A. (1975). Interaction of soluble fibroblast surface antigen with fibrinogen and fibrin: identity with cold insoluble globulin of human plasma. J Exp Med, 141, 497-501.
S.Patel, Ramila, Odermatt, Erich, E.Schwarzbauer, Jean, & O.Hynes, Richard. (1989). Organization of the fibronectin gene provides evidence for exon shuffling during evolution. The EMBO Journal, 6(9), 2565-2572.
Sanjay L. Bhave, Theodoros N. Teknos, and Quintin Pan. (2011). Molecular parameters of head and neck cancer metastasis. Crit Rev Eukaryot Gene Expr., 2011(21), 2.
Skalniak, L., Mizgalska, D., Zarebski, A., Wyrzykowska, P., Koj, A., & Jura, J. (2009). Regulatory feedback loop between NF-kappaB and MCP-1-induced protein 1 RNase. Febs j, 276(20), 5892-5905. doi: 10.1111/j.1742-4658.2009.07273.x
Sturgis, E. M., & Cinciripini, P. M. (2007). Trends in head and neck cancer incidence in relation to smoking prevalence: an emerging epidemic of human papillomavirus-associated cancers? Cancer, 110(7), 1429-1435. doi: 10.1002/cncr.22963
Takagi, M., Absalon, M. J., McLure, K. G., & Kastan, M. B. (2005). Regulation of p53 translation and induction after DNA damage by ribosomal protein L26 and nucleolin. Cell, 123(1), 49-63. doi: 10.1016/j.cell.2005.07.034
Thomas, S. M., Bhola, N. E., Zhang, Q., Contrucci, S. C., Wentzel, A. L., Freilino, M. L., . . . Grandis, J. R. (2006). Cross-talk between G protein-coupled receptor and epidermal growth factor receptor signaling pathways contributes to growth and invasion of head and neck squamous cell carcinoma. Cancer Res, 66(24), 11831-11839. doi: 10.1158/0008-5472.CAN-06-2876
Tsai, N. P., Lin, Y. L., & Wei, L. N. (2009). MicroRNA mir-346 targets the 5'-untranslated region of receptor-interacting protein 140 (RIP140) mRNA and up-regulates its protein expression. Biochem J, 424(3), 411-418. doi: 10.1042/bj20090915
Tsou, J. H., Chang, K. Y., Wang, W. C., Tseng, J. T., Su, W. C., Hung, L. Y., . . . Chen, B. K. (2008). Nucleolin regulates c-Jun/Sp1-dependent transcriptional activation of cPLA2alpha in phorbol ester-treated non-small cell lung cancer A549 cells. Nucleic Acids Res, 36(1), 217-227. doi: 10.1093/nar/gkm1027
Weppelmann B, Wheeler RH, Peters GE, Kim RY, Spencer SA, Meredith RF, Salter MM. (1992). Treatment of recurrent head and neck cancer with 5-fluorouracil, hydroxyurea, and reirradiation. Int J Radiat Oncol Biol Phys., 22(5), 1051-1056.
Wroblewski, L. E., Pritchard, D. M., Carter, S., & Varro, A. (2002). Gastrin-stimulated gastric epithelial cell invasion: the role and mechanism of increased matrix metalloproteinase 9 expression. Biochem J, 365(Pt 3), 873-879. doi: 10.1042/bj20020068
Yang, J., & Weinberg, R. A. (2008). Epithelial-mesenchymal transition: at the crossroads of development and tumor metastasis. Dev Cell, 14(6), 818-829. doi: 10.1016/j.devcel.2008.05.009
Yang TL, Wu TL, Ko TL, Wang TL, Lou TL and Chang TL. (2008). Significance of tumor satellite variables in reflecting the epithelial-mesenchymal transition of tongue cancer. Oral Oncol, 47(8), 720-724.
Yasutaka Moriguchi, Hiroaki Matsubara, Yasukiyo Mori, Satoshi Murasawa, Hiroya Masaki, Katsuya Maruyama, Yoshiaki Tsutsumi, Yasunobu Shibasaki, Yoko Tanaka, Takuma Nakajima, Kinichiro Oda, Toshiji Iwasaka. (1999). Angiotensin II–Induced Transactivation of Epidermal Growth Factor Receptor Regulates Fibronectin and Transforming Growth Factor-b Synthesis via Transcriptional and Posttranscriptional Mechanisms. Circ Res, 84, 1073-1084.
Yousif, N. G. (2014). Fibronectin promotes migration and invasion of ovarian cancer cells through up-regulation of FAK-PI3K/Akt pathway. Cell Biol Int, 38(1), 85-91. doi: 10.1002/cbin.10184
Zimmermann, M., Zouhair, A., Azria, D., & Ozsahin, M. (2006). The epidermal growth factor receptor (EGFR) in head and neck cancer: its role and treatment implications. Radiat Oncol, 1, 11. doi: 10.1186/1748-717X-1-11