Effects of RING-SH2^(Grb2), a Chimeric Protein Containing the E3 Ligase Domain of Cbl, on the EGFR Pathway

10.4077/CJP.2014.BAD281

Wei-Hao Lee；Pei-Yu Wang；Yu-Hung Lin；He-Yen Chou；Yen-Hsien Lee；Chien-Kuo Lee；Li-Mei Pai

c-Cbl ； EGFR ； Grb2 ； lung cancer ； RING domain

The Chinese Journal of Physiology

57卷6期（2014 / 12 / 31）

350 - 357

英文

The E3 ubiquitin-protein ligase Casitas B-lineage lymphoma protein (Cbl) negatively regulates epidermal growth factor receptor (EGFR) signaling pathway in many organisms, and has crucial roles in cell growth, development and human pathologies, including lung cancers. RING-SH2^(Grb2) a chimeric protein of 215 amino acids containing the RING domain of Cbl that provides E3 ligase activity, and t he SH2 domain of Grb2 that serves as an adaptor for EGFR. In this study, we demonstrated that RING-SH2^(Grb2) could promote the ubiquitinylation and degradation of EGFR in a human non-small cell lung carcinoma cell line H1299. Moreover, we discovered that the RING-SH2^(Grb2) chimera promoted the internalization of ligand-bound EGFR, inhibited the growth of H1299 cells, and significantly suppressed tumor growth in a xenograft mouse model. In summary, our results revealed a potential new cancer therapeutic approach for non-small cell lung cancer.

1. Tsai, W.C.,Lee, H.S.,Jin, J.S.,Gao, H.W.,Chao, T.K.,Chen, A.,Nieh, S.,Chan, D.C.,Chang, F.N.,Lin, C.K.(2012).Association between Osteopontin and EGFR expression with clinicopathological parameters in hepatocellular carcinoma.Chinese J. Physiol.,55,412-420.
   連結：
2. Avraham, R.,Yarden, Y.(2011).Feedback regulation of EGFR signalling: decision making by early and delayed loops.Nat. Rev. Mol. Cell Biol.,12,104-117.
3. Chen, Y.R.,Fu, Y.N.,Lin, C.H.,Yang, S.T.,Hu, S.F.,Chen, Y.T.,Tsai, S.F.,Huang, S.F.(2006).Distinctive activation patterns in constitutively active and gefitinib-sensitive EGFR mutants.Oncogene,25,1205-1215.
4. Deshaies, R.J.,Joazeiro, C.A.P.(2009).RING domain E3 ubiquitin ligases.Annu. Rev. Biochem.,78,399-434.
5. Duan, L.,Miura, Y.,Dimri, M.,Majumder, B.,Dodge, I.L.,Reddi, A.L.,Ghosh, A.,Fernandes, N.,Zhou, P.,Mullane-Robinson, K.,Rao, N.,Donoghue, S.,Rogers, R.A.,Bowtell, D.,Naramura, M.,Gu, H.,Band, V.,Band, H.(2003).Cbl-mediated ubiquitinylation is required for lysosomal sorting of epidermal growth factor receptor but is dispensable for endocytosis.J. Biol. Chem.,278,28950-28960.
6. Haglund, K.,Di Fiore, P.P.,Dikic, I.(2003).Distinct monoubiquitin signals in receptor endocytosis.Trends Biochem. Sci.,28,598-604.
7. Haglund, K.,Dikic, I.(2012).The role of ubiquitylation in receptor endocytosis and endosomal sorting.J. Cell Sci.,125,265-275.
8. Hirsch, F.R.,Scagliotti, G.V.,Langer, C.J.,Varella-Garcia, M.,Franklin, W.A.(2003).Epidermal growth factor family of receptors in preneoplasia and lung cancer: perspectives for targeted therapies.Lung Cancer,41(Suppl 1),S29-S42.
9. Huang, F.,Sorkin, A.(2005).Growth factor receptor binding protein 2-mediated recruitment of the RING domain of Cbl to the epidermal growth factor receptor is essential and sufficient to support receptor endocytosis.Mol. Biol. Cell,16,1268-1281.
10. Kiyatkin, A.,Aksamitiene, E.,Markevich, N.I.,Borisov, N.M.,Hoek, J.B.,Kholodenko, B.N.(2006).Scaffolding protein Grb2-associated binder 1 sustains epidermal growth factor-induced mitogenic and survival signaling by multiple positive feedback loops.J. Biol. Chem.,281,19925-19938.
11. Levkowitz, G.,Waterman, H.,Ettenberg, S.A.,Katz, M.,Tsygankov, A.Y.,Alroy, I.,Lavi, S.,Iwai, K.,Reiss, Y.,Ciechanover, A.,Lipkowitz, S.,Yarden, Y.(1999).Ubiquitin ligase activity and tyrosine phosphorylation underlie suppression of growth factor signaling by c-Cbl/Sli-1.Mol. Cell,4,1029-1040.
12. Machida, K.,Mayer, B.J.(2005).The SH2 domain: versatile signaling module and pharmaceutical target.Biochim. Biophys. Acta,1747,1-25.
13. Martarelli, D.,Catalano, A.,Procopio, A.,Orecchia, S.,Libener, R.,Santoni, G.(2006).Characterization of human malignant mesothelioma cell lines orthotopically implanted in the pleural cavity of immunodeficient mice for their ability to grow and form metastasis.BMC Cancer,6,130.
14. Mendelsohn, J.,Baselga, J.(2006).Epidermal growth factor receptor targeting in cancer.Semin. Oncol.,33,369-385.
15. Normanno, N.,De Luca, A.,Bianco, C.,Strizzi, L.,Mancino, M.,Maiello, M.R.,Carotenuto, A.,De Feo, G.,Caponigro, F.,Salomon, D.S.(2006).Epidermal growth factor receptor (EGFR) signaling in cancer.Gene,366,2-16.
16. Polo, S.(2012).Signaling-mediated control of ubiquitin ligases in endocytosis.BMC Biol.,10,25.
17. Rubin, C.,Gur, G.,Yarden, Y.(2005).Negative regulation of receptor tyrosine kinases: unexpected links to c-Cbl and receptor ubiquitylation.Cell Res.,15,66-71.
18. Scaife, R.M.,Langdon, W.Y.(2000).c-Cbl localizes to actin lamellae and regulates lamellipodia formation and cell morphology.J. Cell Sci.,113,215-226.
19. Schmidt, M.H.,Dikic, I.(2005).The Cbl interactome and its functions.Nat. Rev. Mol. Cell Biol.,6,907-918.
20. Sithanandam, G.,Smith, G.T.,Fields, J.R.,Fornwald, L.W.,Anderson, L.M.(2005).Alternate paths from epidermal growth factor receptor to Akt in malignant versus nontransformed lung epithelial cells: ErbB3 versus Gab1.Am. J. Respir. Cell Mol. Biol.,33,490-499.
21. Sorkin, A.,Goh, L.K.(2009).Endocytosis and intracellular trafficking of ErbBs.Exp. Cell Res.,315,683-696.
22. Wang, P.Y.,Pai, L.M.(2011).D-Cbl binding to Drk leads to dose-dependent down-regulation of EGFR signaling and increases receptor-ligand endocytosis.PLoS One,6,e17097.
23. Waterman, H.,Katz, M.,Rubin, C.,Shtiegman, K.,Lavi, S.,Elson, A.,Jovin, T.,Yarden, Y.(2002).A mutant EGF-receptor defective in ubiquitylation and endocytosis unveils a role for Grb2 in negative signaling.EMBO J.,21,303-313.
24. Yarden, Y.,Shilo, B.Z.(2007).SnapShot: EGFR signaling pathway.Cell,131,1018.