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Molecular Cancer
Published in: Molecular Cancer 1/2018

Open Access 01-12-2018 | Research

The E3 ubiquitin ligase NEDD4 mediates cell migration signaling of EGFR in lung cancer cells

Authors: Genbao Shao, Ranran Wang, Aiqin Sun, Jing Wei, Ke Peng, Qian Dai, Wannian Yang, Qiong Lin

Published in: Molecular Cancer | Issue 1/2018

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Abstract

Background

EGFR-dependent cell migration plays an important role in lung cancer progression. Our previous study observed that the HECT E3 ubiquitin ligase NEDD4 is significantly correlated with tumor metastasis and required for migration and invasion signaling of EGFR in gastric cancer cells. However, how NEDD4 promotes the EGFR-dependent lung cancer cell migration is unknown. This study is to elucidate the mechanism by which NEDD4 mediates the EGFR lung cancer migration signaling.

Methods

Lentiviral vector-loaded NEDD4 shRNA was used to deplete endogenous NEDD4 in lung cancer cell lines. Effects of the NEDD4 knockdown on the EGFR-dependent or independent lung cancer cell migration were determined using the wound-healing and transwell assays. Association of NEDD4 with activated EGFR was assayed by co-immunoprecipitation. Co-expression of NEDD4 with EGFR or PTEN was determined by immunohistochemical (IHC) staining in 63 lung adenocarcinoma tissue samples. Effects of NEDD4 ectopic expression or knockdown on PTEN ubiquitination and down-regulation, AKT activation and lysosomal secretion were examined using the GST-Uba pulldown assay, immunoblotting, immunofluorescent staining and a human cathepsin B ELISA assay respectively. The specific cathepsin B inhibitor CA-074Me was used for assessing the role of cathepsin B in lung cancer cell migration.

Results

Knockdown of NEDD4 significantly reduced EGF-stimulated cell migration in non-small cell lung carcinoma (NSCLC) cells. Co-immunoprecipitation assay found that NEDD4 is associated with EGFR complex upon EGF stimulation, and IHC staining indicates that NEDD4 is co-expressed with EGFR in lung adenocarcinoma tumor tissues, suggesting that NEDD4 might mediate lung cancer cell migration by interaction with the EGFR signaling complex. Interestingly, NEDD4 promotes the EGF-induced cathepsin B secretion, possibly through lysosomal exocytosis, as overexpression of the ligase-dead mutant of NEDD4 impedes lysosomal secretion, and knockdown of NEDD4 significantly reduced extracellular amount of cathepsin B induced by EGF. Consistent with the role of NEDD4, cathepsin B is pivotal for both basal and the EGF-stimulated lung cancer cell migration. Our studies propose a novel mechanism underlying the EGFR-promoted lung cancer cell migration that is mediated by NEDD4 through regulation of cathepsin B secretion.

Conclusion

NEDD4 mediates the EGFR lung cancer cell migration signaling through promoting lysosomal secretion of cathepsin B.
Literature
1.
Staub O, Gautschi I, Ishikawa T, Breitschopf K, Ciechanover A, Schild L, Rotin D. Regulation of stability and function of the epithelial Na+ channel (ENaC) by ubiquitination. EMBO J. 1997;16(21):6325–36.CrossRefPubMedPubMedCentral
2.
Staub O, Dho S, Henry P, Correa J, Ishikawa T, McGlade J, Rotin DWW. Domains of Nedd4 bind to the proline-rich PY motifs in the epithelial Na+ channel deleted in Liddle's syndrome. EMBO J. 1996;15(10):2371–80.PubMedPubMedCentral
3.
Persaud A, Alberts P, Amsen EM, Xiong X, Wasmuth J, Saadon Z, Fladd C, Parkinson J, Rotin D. Comparison of substrate specificity of the ubiquitin ligases Nedd4 and Nedd4-2 using proteome arrays. Mol Syst Biol. 2009;5:333.CrossRefPubMedPubMedCentral
4.
Boase NA, Kumar S. NEDD4: the founding member of a family of ubiquitin-protein ligases. Gene. 2015;557(2):113–22.CrossRefPubMed
5.
Ren J, Kee Y, Huibregtse JM, Piper RC. Hse1, a component of the yeast Hrs-STAM ubiquitin-sorting complex, associates with ubiquitin peptidases and a ligase to control sorting efficiency into multivesicular bodies. Mol Biol Cell. 2007;18(1):324–35.CrossRefPubMedPubMedCentral
6.
Léon S, Erpapazoglou Z, Haguenauer-Tsapis R. Ear1p and Ssh4p are new adaptors of the ubiquitin ligase Rsp5p for cargo ubiquitylation and sorting at multivesicular bodies. Mol Biol Cell. 2008;19(6):2379–88.CrossRefPubMedPubMedCentral
7.
Magnifico A, Ettenberg S, Yang C, Mariano J, Tiwari S, Fang S, Lipkowitz S, Weissman AM. WW domain HECT E3s target Cbl RING finger E3s for proteasomal degradation. J Biol Chem. 2003;278:43169–77.CrossRefPubMed
8.
Katz M, Shtiegman K, Tal-Or P, Yakir L, Mosesson Y, Harari D, Machluf Y, Asao H, Jovin T, Sugamura K, Yarden Y. Ligand-independent degradation of epidermal growth factor receptor involves receptor ubiquitylation and Hgs, an adaptor whose ubiquitin-interacting motif targets ubiquitylation by Nedd4. Traffic. 2002;3(10):740–51.CrossRefPubMed
9.
Segura-Morales C, Pescia C, Chatellard-Causse C, Sadoul R, Bertrand E, Basyuk E. Tsg101 and Alix interact with murine leukemia virus gag and cooperate with Nedd4 ubiquitin ligases during budding. J Biol Chem. 2005;280(29):27004–12.CrossRefPubMed
10.
Woelk T, Oldrini B, Maspero E, Confalonieri S, Cavallaro E, Di Fiore PP, Polo S. Molecular mechanisms of coupled monoubiquitination. Nat Cell Biol. 2006;8:1246–54.CrossRefPubMed
11.
Blot V, Perugi F, Gay B, Prévost MC, Briant L, Tangy F, Abriel H, Staub O, Dokhélar MC, Pique C. Nedd4.1-mediated ubiquitination and subsequent recruitment of Tsg101 ensure HTLV-1 gag trafficking towards the multivesicular body pathway prior to virus budding. J Cell Sci. 2004;117(Pt 11):2357–67.CrossRefPubMed
12.
Aoh QL, Castle AM, Hubbard CH, Katsumata O, Castle JD. SCAMP3 negatively regulates epidermal growth factor receptor degradation and promotes receptor recycling. Mol Biol Cell. 2009;20(6):1816–32.CrossRefPubMedPubMedCentral
13.
Lin Q, Wang J, Childress C, Sudol M, Carey DJ, Yang WHECT. E3 ubiquitin ligase Nedd4-1 ubiquitinates ACK and regulates EGF-induced degradation of EGFR and ACK. Mol Cell Biol. 2010;30:1541–54.CrossRefPubMedPubMedCentral
14.
Sun A, Wei J, Childress C, Shaw JH 4th, Peng K, Shao G, Yang W, Lin Q. The E3 ubiquitin ligase NEDD4 is an LC3-interactive protein and regulates autophagy. Autophagy 2017, 13(3):522–537.
15.
Lin Q, Dai Q, Meng H, Sun A, Wei J, Peng K, Childress C, Chen M, Shao G, Yang W. The HECT E3 ubiquitin ligase NEDD4 interacts with and ubiquitylates SQSTM1 for inclusion body autophagy. J Cell Sci. 2017;130(22):3839–50.CrossRefPubMed
16.
Zou X, Levy-Cohen G, Blank M. Molecular functions of NEDD4 E3 ubiquitin ligases in cancer. Biochim Biophys Acta. 2015;1856(1):91–106.PubMed
17.
Wang X, Trotman LC, Koppie T, Alimonti A, Chen Z, Gao Z, Wang J, Erdjument-Bromage H, Tempst P, Cordon-Cardo C, Pandolfi PP, Jiang X. NEDD4-1 is a proto-oncogenic ubiquitin ligase for PTEN. Cell. 2007;128:129–39.CrossRefPubMedPubMedCentral
18.
Trotman LC, Wang X, Alimonti A, Chen Z, Teruya-Feldstein J, Yang H, Pavletich NP, Carver BS, Cordon-Cardo C, Erdjument-Bromage H, Tempst P, Chi SG, Kim HJ, Misteli T, Jiang X, Pandolfi PP. Ubiquitination regulates PTEN nuclear import and tumor suppression. Cell. 2007;128(1):141–56.CrossRefPubMedPubMedCentral
19.
Shen WH, Balajee AS, Wang J, Wu H, Eng C, Pandolfi PP, Yin Y. Essential role for nuclear PTEN in maintaining chromosomal integrity. Cell. 2007;128(1):157–70.CrossRefPubMed
20.
Sun A, Yu G, Dou X, Yan X, Yang W, Lin Q. Nedd4-1 is an exceptional prognostic biomarker for gastric cardia adenocarcinoma and functionally associated with metastasis. Mol Cancer. 2014;13:248.CrossRefPubMedPubMedCentral
21.
Baselga J, Albanell J. Targeting epidermal growth factor receptor in lung cancer. Curr Oncol Rep. 2002;4(4):317–24.CrossRefPubMed
22.
Arteaga CL, Johnson DH. Tyrosine kinase inhibitors-ZD1839 (Iressa). Curr Opin Oncol. 2001;13(6):491–8.CrossRefPubMed
23.
Herbst RS, Kies MS. ZD1839 (Iressa) in non-small cell lung cancer. Oncologist. 2002;7(Suppl 4):9–15.CrossRefPubMed
24.
Khazaie K, Schirrmacher V, Lichtner RB. EGF receptor in neoplasia and metastasis. Cancer Metastasis Rev. 1993;12(3–4):255–74.CrossRefPubMed
25.
Damstrup L, Rude Voldborg B, Spang-Thomsen M, Brünner N, Skovgaard Poulsen H. In vitro invasion of small-cell lung cancer cell lines correlates with expression of epidermal growth factor receptor. Br J Cancer. 1998;78(5):631–40.CrossRefPubMedPubMedCentral
26.
Ritter CA, Arteaga CL. The epidermal growth factor receptor-tyrosine kinase: a promising therapeutic target in solid tumors. Semin Oncol. 2003;30(1 Suppl 1):3–11.CrossRefPubMed
27.
Nicholson RI, Gee JM, Harper ME. EGFR and cancer prognosis. Eur J Cancer. 2001;37(Suppl 4):S9–15.CrossRefPubMed
28.
Marcoux N, Vuori K. EGF receptor mediates adhesion-dependent activation of the Rac GTPase: a role for phosphatidylinositol 3-kinase and Vav2. Oncogene. 2003;22(38):6100–6.CrossRefPubMed
29.
Chung BM, Dimri M, George M, Reddi AL, Chen G, Band V, Band H. The role of cooperativity with Src in oncogenic transformation mediated by non-small cell lung cancer-associated EGF receptor mutants. Oncogene. 2009;28(16):1821–32.CrossRefPubMedPubMedCentral
30.
Prenzel N, Fischer OM, Streit S, Hart S, Ullrich A. The epidermal growth factor receptor family as a central element for cellular signal transduction and diversification. Endocr Relat Cancer. 2001;8(1):11–31.CrossRefPubMed
31.
Schäfer B, Gschwind A, Ullrich A. Multiple G-protein-coupled receptor signals converge on the epidermal growth factor receptor to promote migration and invasion. Oncogene. 2004;23(4):991–9.CrossRefPubMed
32.
Chen F, Deng J, Liu X, Li W, Zheng J. HCRP-1 regulates cell migration and invasion via EGFR-ERK mediated up-regulation of MMP-2 with prognostic significance in human renal cell carcinoma. Sci Rep. 2015;5:13470.CrossRefPubMedPubMedCentral
33.
Verma N, Keinan O, Selitrennik M, Karn T, Filipits M, Lev S. PYK2 sustains endosomal-derived receptor signalling and enhances epithelial-to-mesenchymal transition. Nat Commun. 2015;6:6064.CrossRefPubMed
34.
Wang J, Peng Q, Lin Q, Childress C, Carey D, Yang W. Calcium activates Nedd4 E3 ubiquitin ligases by releasing the C2 domain-mediated auto-inhibition. J Biol Chem. 2010;285(16):12279–88.CrossRefPubMedPubMedCentral
35.
Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, Harris PL, Haserlat SM, Supko JG, Haluska FG, Louis DN, Christiani DC, Settleman J, Haber DA. Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med. 2004;350(21):2129–39.CrossRefPubMed
36.
Paez JG, Jänne PA, Lee JC, Tracy S, Greulich H, Gabriel S, Herman P, Kaye FJ, Lindeman N, Boggon TJ, Naoki K, Sasaki H, Fujii Y, Eck MJ, Sellers WR, Johnson BE, Meyerson M. EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy. Science. 2004;304(5676):1497–500.CrossRefPubMed
37.
Sordella R, Bell DW, Haber DA, Settleman J. Gefitinib-sensitizing EGFR mutations in lung cancer activate anti-apoptotic pathways. Science. 2004;305(5687):1163–7.CrossRefPubMed
38.
Shen F, Lin Q, Gu Y, Childress C, Yang W. Activated Cdc42-associated kinase 1 is a component of EGF receptor signaling complex and regulates EGF receptor degradation. Mol Biol Cell. 2007;18(3):732–42.CrossRefPubMedPubMedCentral
39.
Amodio N, Scrima M, Palaia L, Salman AN, Quintiero A, Franco R, Botti G, Pirozzi P, Rocco G, De Rosa N, Viglietto G. Oncogenic role of the E3 ubiquitin ligase NEDD4-1, a PTEN negative regulator, in non-small-cell lung carcinomas. Am J Pathol. 2010;177(5):2622–34.CrossRefPubMedPubMedCentral
40.
Yim EK, Peng G, Dai H, Hu R, Li K, Lu Y, Mills GB, Meric-Bernstam F, Hennessy BT, Craven RJ, Lin SY. Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Cancer Cell. 2009;15(4):304–14.CrossRefPubMedPubMedCentral
41.
Hong SW, Moon JH, Kim JS, Shin JS, Jung KA, Lee WK, Jeong SY, Hwang JJ, Lee SJ, Suh YA, Kim I, Nam KY, Han S, Kim JE, Kim KP, Hong YS, Lee JL, Lee WJ, Choi EK, Lee JS, Jin DH, Kim TW. p34 is a novel regulator of the oncogenic behavior of NEDD4-1 and PTEN. Cell Death Differ. 2014;21(1):146–60.CrossRefPubMed
42.
Marte BM, Downward J. PKB/Akt: connecting phosphoinositide 3-kinase to cell survival and beyond. Trends Biochem Sci. 1997;22(9):355–8.CrossRefPubMed
43.
Kim D, Kim S, Koh H, et al. Akt/PKB promotes cancer cell invasion via increased motility and metalloproteinase production. FASEB J. 2001;15:1953–62.CrossRefPubMed
44.
Fouladkou F, Landry T, Kawabe H, Neeb A, Lu C, Brose N, Stambolic V, Rotin D. The ubiquitin ligase Nedd4-1 is dispensable for the regulation of PTEN stability and localization. Proc Natl Acad Sci U S A. 2008;105(25):8585–90.CrossRefPubMedPubMedCentral
45.
Maddika S, Kavela S, Rani N, Palicharla VR, Pokorny JL, Sarkaria JN, Chen J. WWP2 is an E3 ubiquitin ligase for PTEN. Nat Cell Biol. 2011;13(6):728–33.CrossRefPubMedPubMedCentral
46.
Frey MR, Dise RS, Edelblum KL, Polk DB. p38 kinase regulates epidermal growth factor receptor downregulation and cellular migration. EMBO J. 2006;25(24):5683–92.CrossRefPubMedPubMedCentral
47.
Keppler D, Abrahamson M, Sordat B. Secretion of cathepsin B and tumour invasion. Biochem Soc Trans. 1994;22(1):43–9.CrossRefPubMed
48.
Warren L. Stimulation of lysosomal enzyme secretion by growth factors. Exp Cell Res. 1990;190(1):133–6.CrossRefPubMed
49.
Sung BH, Zhu X, Kaverina I, Weaver AM. Cortactin controls cell motility and lamellipodial dynamics by regulating ECM secretion. Curr Biol. 2011;21(17):1460–9.CrossRefPubMedPubMedCentral
50.
Mi S, Qin XW, Lin YF, He J, Chen NN, Liu C, Weng SP, He JG, Guo CJ. Budding of Tiger frog virus (an Iridovirus) from HepG2 cells via three ways recruits the ESCRT pathway. Sci Rep. 2016;6:26581.CrossRefPubMedPubMedCentral
51.
Harty RN, Brown ME, Wang G, Huibregtse J, Hayes FP. A PPxY motif within the VP40 protein of Ebola virus interacts physically and functionally with a ubiquitin ligase: implications for filovirus budding. Proc Natl Acad Sci U S A. 2000;97:13871–6.CrossRefPubMedPubMedCentral
52.
53.
Sloane BF, Honn KV. Cysteine proteinases and metastasis. Cancer Metastasis Rev. 1984;3(3):249–63.CrossRefPubMed
54.
Podgorski I, Sloane BF. Cathepsin B and its role(s) in cancer progression. Biochem Soc Symp. 2003;70:263–76.CrossRef
55.
Buck MR, Karustis DG, Day NA, Honn KV, Sloane BF. Degradation of extracellular-matrix proteins by human cathepsin B from normal and tumour tissues. Biochem J. 1992;282(Pt 1):273–8.CrossRefPubMedPubMedCentral
56.
Garcia-Cattaneo A, Gobert FX, Müller M, Toscano F, Flores M, Lescure A, Del Nery E, Benaroch P. Cleavage of toll-like receptor 3 by cathepsins B and H is essential for signaling. Proc Natl Acad Sci U S A. 2012;109(23):9053–8.CrossRefPubMedPubMedCentral
57.
Kobayashi H, Schmitt M, Goretzki L, Chucholowski N, Calvete J, Kramer M, Günzler WA, Jänicke F, Graeff H, Cathepsin B. Efficiently activates the soluble and the tumor cell receptor-bound form of the proenzyme urokinase-type plasminogen activator (pro-uPA). J Biol Chem. 1991;266(8):5147–52.PubMed
58.
Tu C, Ortega-Cava CF, Chen G, Fernandes ND, Cavallo-Medved D, Sloane BF, Band V, Band H. Lysosomal cathepsin B participates in the podosome-mediated extracellular matrix degradation and invasion via secreted lysosomes in v-Src fibroblasts. Cancer Res. 2008;68(22):9147–56.CrossRefPubMedPubMedCentral
59.
Schraufstatter IU, Trieu K, Zhao M, Rose DM, Terkeltaub RA, Burger M. IL-8-mediated cell migration in endothelial cells depends on cathepsin B activity and transactivation of the epidermal growth factor receptor. J Immunol. 2003;171(12):6714–22.CrossRefPubMed
60.
Wan X, Helman LJ. Levels of PTEN protein modulate Akt phosphorylation on serine 473, but not on threonine 308, in IGF-II-overexpressing rhabdomyosarcomas cells. Oncogene. 2003;22(50):8205–11.CrossRefPubMed
61.
Huang ZJ, Zhu JJ, Yang XY, Biskup E. NEDD4 promotes cell growth and migration via PTEN/PI3K/AKT signaling in hepatocellular carcinoma. Oncol Lett. 2017;14(3):2649–56.CrossRefPubMedPubMedCentral
62.
Hsia HE, Kumar R, Luca R, Takeda M, Courchet J, Nakashima J, Wu S, Goebbels S, An W, Eickholt BJ, Polleux F, Rotin D, Wu H, Rossner MJ, Bagni C, Rhee JS, Brose N, Kawabe H. Ubiquitin E3 ligase Nedd4-1 acts as a downstream target of PI3K/PTEN-mTORC1 signaling to promote neurite growth. Proc Natl Acad Sci U S A. 2014;111(36):13205–10.CrossRefPubMedPubMedCentral
63.
Manjithaya R, Subramani S. Autophagy: a broad role in unconventional protein secretion? Trends Cell Biol. 2011;21(2):67–73.CrossRefPubMed
Metadata
Title
The E3 ubiquitin ligase NEDD4 mediates cell migration signaling of EGFR in lung cancer cells
Authors
Genbao Shao
Ranran Wang
Aiqin Sun
Jing Wei
Ke Peng
Qian Dai
Wannian Yang
Qiong Lin
Publication date
01-12-2018
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2018
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/s12943-018-0784-2

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