HECTD1 regulates the expression of SNAIL: Implications for epithelial‑mesenchymal transition

Wang,Xinggang; De Geyter,Christian; Jia,Zanhui; Peng,Ya; Zhang,Hong

doi:10.3892/ijo.2020.5002

HECTD1 regulates the expression of SNAIL: Implications for epithelial‑mesenchymal transition

Authors:
- Xinggang Wang
- Christian De Geyter
- Zanhui Jia
- Ya Peng
- Hong Zhang
View Affiliations

Affiliations: Department of Biomedicine (DBM), University Hospital, University of Basel, CH‑4031 Basel, Switzerland, Second Hospital of Jilin University, Changchun, Jilin 130041, P.R. China
Published online on: February 27, 2020 https://doi.org/10.3892/ijo.2020.5002
Pages: 1186-1198
Copyright: © Wang 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

As a transcription factor, SNAIL plays a crucial role in embryonic development and cancer progression by mediating epithelial‑mesenchymal transition (EMT); however, post‑translational modifications, such as ubiquitination, which control the degradation of SNAIL have been observed to affect its functional role in EMT. In a previous study by the authors, it was demonstrated that the HECT domain E3 ubiquitin ligase 1 (HECTD1) regulated the dynamic nature of adhesive structures. In the present study, HECTD1 was observed to interact with SNAIL and regulate its stability through ubiquitination, and the knockdown of HECTD1 increased the expression levels of SNAIL. HECTD1 was discovered to contain putative nuclear localization and export signals that facilitated its translocation between the cytoplasm and nucleus, a process regulated by epidermal growth factor (EGF). Treatment with leptomycin B resulted in the nuclear retention of HECTD1, which was associated with the loss of SNAIL expression. The knockdown of HECTD1 in HeLa cells increased cell migration and induced a mesenchymal phenotype, in addition to demonstrating sustained EGF signaling, which was observed through increased phosphorylated ERK expression levels. Under hypoxic conditions, HECTD1 expression levels were decreased by microRNA (miRNA or miR)‑210. Upon the observation of genetic abnormalities in the HECTD1 gene in cervical cancer specimens, it was observed that the decreased expression levels of HECTD1 were significantly associated with a poor patient survival. Thus, it was hypothesized that HECTD1 may regulate EMT through the hypoxia/hypoxia inducible factor 1α/miR‑210/HECTD1/SNAIL signaling pathway and the EGF/EGF receptor/HECTD1/ERK/SNAIL signaling pathway in cervical cancer. On the whole, the data of the present study indicated that HECTD1 serves as an E3 ubiquitin ligase to mediate the stability of SNAIL proteins.

View Figures

View References

1	Sahu SK, Tiwari N, Pataskar A, Zhuang Y, Borisova M, Diken M, Strand S, Beli P and Tiwari VK: FBXO32 promotes microenvironment underlying epithelial-mesenchymal transition via CtBP1 during tumour metastasis and brain development. Nat Commun. 8:15232017. View Article : Google Scholar : PubMed/NCBI
2	Liu Y, Zhou H, Zhu R, Ding F, Li Y, Cao X and Liu Z: SPSB3 targets SNAIL for degradation in GSK-3beta phosphorylation-dependent manner and regulates metastasis. Oncogene. 37:768–776. 2018. View Article : Google Scholar
3	Park SH, Jung EH, Kim GY, Kim BC, Lim JH and Woo CH: Itch E3 ubiquitin ligase positively regulates TGF-β signaling to EMT via smad7 ubiquitination. Mol Cells. 38:20–25. 2015. View Article : Google Scholar
4	Van Laar RK: Genomic signatures for predicting survival and adjuvant chemotherapy benefit in patients with non-small-cell lung cancer. BMC Med Genomics. 5:302012. View Article : Google Scholar : PubMed/NCBI
5	Lim SK, Lu SY, Kang SA, Tan HJ, Li Z, Adrian Wee ZN, Guan JS, Reddy Chichili VP, Sivaraman J, Putti T, et al: Wnt signaling promotes breast cancer by blocking ITCH-Mediated degradation of YAP/TAZ transcriptional coactivator WBP2. Cancer Res. 76:6278–6289. 2016. View Article : Google Scholar : PubMed/NCBI
6	Zhou H, Liu Y, Zhu R, Ding F, Wan Y, Li Y and Liu Z: FBXO32 suppresses breast cancer tumorigenesis through targeting KLF4 to proteasomal degradation. Oncogene. 36:3312–3321. 2017. View Article : Google Scholar : PubMed/NCBI
7	Komander D: The emerging complexity of protein ubiquitination. Biochem Soc Trans. 37:937–953. 2009. View Article : Google Scholar : PubMed/NCBI
8	Pickart CM and Eddins MJ: Ubiquitin: Structures, functions, mechanisms. Biochim Biophys Acta. 1695:55–72. 2004. View Article : Google Scholar : PubMed/NCBI
9	Metzger MB, Hristova VA and Weissman AM: HECT and RING finger families of E3 ubiquitin ligases at a glance. J Cell Sci. 125:531–537. 2012. View Article : Google Scholar : PubMed/NCBI
10	Scheffner M and Kumar S: Mammalian HECT ubiquitin-protein ligases: Biological and pathophysiological aspects. Biochim Biophys Acta. 1843:61–74. 2014. View Article : Google Scholar
11	D'Alonzo D, Emch FH, Shen X, Bruder E, De Geyter C and Zhang H: Hectd1 is essential for embryogenesis in mice. Gene Expr Patterns. 10:1190642019. View Article : Google Scholar
12	Shen X, Jia Z, D'Alonzo D, Wang X, Bruder E, Emch FH, De Geyter C and Zhang H: HECTD1 controls the protein level of IQGAP1 to regulate the dynamics of adhesive structures. Cell Commun Signal. 15:22017. View Article : Google Scholar : PubMed/NCBI
13	Zhu S, Si ML, Wu H and Mo YY: MicroRNA-21 targets the tumor suppressor gene tropomyosin 1 (TPM1). J Biol Chem. 282:14328–14336. 2007. View Article : Google Scholar : PubMed/NCBI
14	Besnier M, Gasparino S, Vono R, Sangalli E, Facoetti A, Bollati V, Cantone L, Zaccagnini G, Maimone B, Fuschi P, et al: MiR-210 enhances the therapeutic potential of bone-marrow-derived circulating proangiogenic cells in the setting of limb ischemia. Mol Ther. 26:1694–1705. 2018. View Article : Google Scholar : PubMed/NCBI
15	Fan Q, Hu X, Zhang H, Wang S, Zhang H, You C, Zhang CY, Liang H, Chen X and Ba Y: MiR-193a-3p is an important tumour suppressor in lung cancer and directly targets KRAS. Cell Physiol Biochem. 44:1311–1324. 2017. View Article : Google Scholar : PubMed/NCBI
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
17	Kaundal R and Raghava GP: RSLpred: An integrative system for predicting subcellular localization of rice proteins combining compositional and evolutionary information. Proteomics. 9:2324–2342. 2009. View Article : Google Scholar : PubMed/NCBI
18	Nakai K and Horton P: PSORT: A program for detecting sorting signals in proteins and predicting their subcellular localization. Trends Biochem Sci. 24:34–36. 1999. View Article : Google Scholar : PubMed/NCBI
19	Vinas-Castells R, Frias A, Robles-Lanuza E, Zhang K, Longmore GD, García de Herreros A and Díaz VM: Nuclear ubiquitination by FBXL5 modulates Snail1 DNA binding and stability. Nucleic Acids Res. 42:1079–1094. 2014. View Article : Google Scholar :
20	Zheng H, Shen M, Zha YL, Li W, Wei Y, Blanco MA, Ren G, Zhou T, Storz P, Wang HY and Kang Y: PKD1 phosphory-lation-dependent degradation of SNAIL by SCF-FBXO11 regulates epithelial-mesenchymal transition and metastasis. Cancer Cell. 26:358–373. 2014. View Article : Google Scholar : PubMed/NCBI
21	Zhou BP, Deng J, Xia W, Xu J, Li YM, Gunduz M and Hung MC: Dual regulation of snail by GSK-3beta-mediated phosphorylation in control of epithelial-mesenchymal transition. Nat Cell Biol. 6:931–940. 2004. View Article : Google Scholar : PubMed/NCBI
22	Hutten S and Kehlenbach RH: CRM1-mediated nuclear export: To the pore and beyond. Trends Cell Biol. 17:193–201. 2007. View Article : Google Scholar : PubMed/NCBI
23	Wolff B, Sanglier JJ and Wang Y: Leptomycin B is an inhibitor of nuclear export: Inhibition of nucleo-cytoplasmic translocation of the human immunodeficiency virus type 1 (HIV-1) Rev protein and Rev-dependent mRNA. Chem Biol. 4:139–147. 1997. View Article : Google Scholar : PubMed/NCBI
24	Chaudhry P, Fabi F, Singh M, Parent S, Leblanc V and Asselin E: Prostate apoptosis response-4 mediates TGF-β-induced epithelial-to-mesenchymal transition. Cell Death Dis. 5:e10442014. View Article : Google Scholar
25	Schrevel M, Gorter A, Kolkman-Uljee SM, Trimbos JB, Fleuren GJ and Jordanova ES: Molecular mechanisms of epidermal growth factor receptor overexpression in patients with cervical cancer. Mod Pathol. 24:720–728. 2011. View Article : Google Scholar : PubMed/NCBI
26	Kusewitt DF, Choi C, Newkirk KM, Leroy P, Li Y, Chavez MG and Hudson LG: Slug/Snai2 is a downstream mediator of epidermal growth factor receptor-stimulated reepithelialization. J Invest Dermatol. 129:491–495. 2009. View Article : Google Scholar
27	Zeisberg M and Neilson EG: Biomarkers for epithelial-mesen-chymal transitions. J Clin Invest. 119:1429–1437. 2009. View Article : Google Scholar : PubMed/NCBI
28	Lee MY, Chou CY, Tang MJ and Shen MR: Epithelial-Mesenchymal transition in cervical cancer: Correlation with tumor progression, epidermal growth factor receptor overexpression, and snail up-regulation. Clin Cancer Res. 14:4743–4750. 2008. View Article : Google Scholar : PubMed/NCBI
29	Gan Y, Shi C, Inge L, Hibner M, Balducci J and Huang Y: Differential roles of ERK and Akt pathways in regulation of EGFR-mediated signaling and motility in prostate cancer cells. Oncogene. 29:4947–4958. 2010. View Article : Google Scholar : PubMed/NCBI
30	Misra A, Pandey C, Sze SK and Thanabalu T: Hypoxia activated EGFR signaling induces epithelial to mesenchymal transition (EMT). PLoS One. 7:e497662012. View Article : Google Scholar : PubMed/NCBI
31	Lamouille S, Xu J and Derynck R: Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol. 15:178–196. 2014. View Article : Google Scholar : PubMed/NCBI
32	Zavadil J, Narasimhan M, Blumenberg M and Schneider RJ: Transforming growth factor-beta and microRNA:MRNA regulatory networks in epithelial plasticity. Cells Tissues Organs. 185:157–161. 2007. View Article : Google Scholar : PubMed/NCBI
33	Ding L, Zhao L, Chen W, Liu T, Li Z and Li X: MiR-210, a modulator of hypoxia-induced epithelial-mesenchymal transition in ovarian cancer cell. Int J Clin Exp Med. 8:2299–2307. 2015.PubMed/NCBI
34	Fasanaro P, Greco S, Lorenzi M, Pescatori M, Brioschi M, Kulshreshtha R, Banfi C, Stubbs A, Calin GA and Ivan M: An integrated approach for experimental target identification of hypoxia-induced miR-210. J Biol Chem. 284:35134–35143. 2009. View Article : Google Scholar : PubMed/NCBI
35	Cicchillitti L, Di Stefano V, Isaia E, Crimaldi L, Fasanaro P, Ambrosino V, Antonini A, Capogrossi MC, Gaetano C, Piaggio G and Martelli F: Hypoxia-Inducible factor 1-α induces miR-210 in normoxic differentiating myoblasts. J Biol Chem. 287:44761–44771. 2012. View Article : Google Scholar : PubMed/NCBI
36	Forbes SA, Tang G, Bindal N, Bamford S, Dawson E, Cole C, Kok CY, Jia M, Ewing R, Menzies A, et al: COSMIC (the Catalogue of Somatic Mutations in Cancer): A resource to investigate acquired mutations in human cancer. Nucleic Acids Res. 38:D652–D657. 2010. View Article : Google Scholar :
37	Tomczak K, Czerwinska P and Wiznerowicz M: The cancer genome atlas (TCGA): An immeasurable source of knowledge. Contemp Oncol (Pozn). 19:A68–A77. 2015.
38	Uhlen M, Zhang C, Lee S, Sjöstedt E, Fagerberg L, Bidkhori G, Benfeitas R, Arif M, Liu Z, Edfors F, et al: A pathology atlas of the human cancer transcriptome. Science. 357:63522017. View Article : Google Scholar
39	Lanczky A, Nagy A, Bottai G, Munkácsy G, Szabó A, Santarpia L and Győrffy B: MiRpower: A web-tool to validate survival-associated miRNAs utilizing expression data from 2178 breast cancer patients. Breast Cancer Res Treat. 160:439–446. 2016. View Article : Google Scholar : PubMed/NCBI
40	Rao Q, Shen Q, Zhou H, Peng Y, Li J and Lin Z: Aberrant microRNA expression in human cervical carcinomas. Med Oncol. 29:1242–1248. 2012. View Article : Google Scholar
41	Zohn IE, Anderson KV and Niswander L: The hectd1 ubiquitin ligase is required for development of the head mesenchyme and neural tube closure. Dev Biol. 306:208–221. 2007. View Article : Google Scholar : PubMed/NCBI
42	Hugel S, Depping R, Dittmar G, Rother F, Cabot R, Sury MD, Hartmann E and Bader M: Identification of importin α 7 specific transport cargoes using a proteomic screening approach. Mol Cell Proteomics. 13:1286–1298. 2014. View Article : Google Scholar
43	Zada S, Hwang JS, Ahmed M, Lai TH, Pham TM and Kim DR: Control of the epithelial-to-mesenchymal transition and cancer metastasis by autophagy-dependent SNAI1 Degradation. Cells. 8:E1292019. View Article : Google Scholar : PubMed/NCBI
44	Liu G, Rogers J, Murphy CT and Rongo C: EGF signalling activates the ubiquitin proteasome system to modulate C. elegans lifespan. EMBO J. 30:2990–3003. 2011. View Article : Google Scholar : PubMed/NCBI
45	Argenzio E, Bange T, Oldrini B, Bianchi F, Peesari R, Mari S, Di Fiore PP, Mann M and Polo S: Proteomic snapshot of the EGF-induced ubiquitin network. Mol Syst Biol. 7:4622011. View Article : Google Scholar : PubMed/NCBI
46	Piret JP, Mottet D, Raes M and Michiels C: CoCl2, a chemical inducer of hypoxia-inducible factor-1, and hypoxia reduce apop-totic cell death in hepatoma cell line HepG2. Annals N Y Acad Sci. 973:443–447. 2002. View Article : Google Scholar
47	Djidja MC, Chang J, Hadjiprocopis A, Schmich F, Sinclair J, Mršnik M, Schoof EM, Barker HE, Linding R, Jørgensen C and Erler JT: Identification of hypoxia-regulated proteins using MALDI-mass spectrometry imaging combined with quantitative proteomics. J Proteome Res. 13:2297–2313. 2014. View Article : Google Scholar : PubMed/NCBI
48	Lester RD, Jo M, Montel V, Takimoto S and Gonias SL: UPAR induces epithelial-mesenchymal transition in hypoxic breast cancer cells. J Cell Biol. 178:425–436. 2007. View Article : Google Scholar : PubMed/NCBI
49	Duhamel S, Goyette MA, Thibault MP, Filion D, Gaboury L and Cote JF: The E3 ubiquitin ligase HectD1 suppresses EMT and metastasis by targeting the +TIP ACF7 for degradation. Cell Rep. 22:1016–1030. 2018. View Article : Google Scholar : PubMed/NCBI