Muscleblind‑like 1 destabilizes Snail mRNA and suppresses the metastasis of colorectal cancer cells via the Snail/E‑cadherin axis

Tang,Liang; Zhao,Peng; Kong,Dalu

doi:10.3892/ijo.2019.4691

Muscleblind‑like 1 destabilizes Snail mRNA and suppresses the metastasis of colorectal cancer cells via the Snail/E‑cadherin axis

Authors:
- Liang Tang
- Peng Zhao
- Dalu Kong
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Affiliations: Department of Colorectal Cancer, Cancer Hospital of Tianjin Medical University, Key Laboratory of Cancer Prevention and Therapy, and National Clinical Research Center of Cancer, Tianjin 300060, P.R. China
Published online on: January 18, 2019 https://doi.org/10.3892/ijo.2019.4691
Pages: 955-965
Copyright: © Tang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

RNA‑binding proteins (RBPs) play a fundamental role in the recurrence and metastasis of colorectal cancer (CRC). In this study, we identified muscleblind‑like 1 (MBNL1), an RBP implicated in developmental control, as a robust suppressor of CRC cell metastasis in vitro. By using a scratch assay coupled with time‑lapse live cell imaging, our findings revealed that the knockdown of MBNL1 induced epithelial‑to‑mesenchymal transition (EMT)‑like morphological changes in the HCT‑116 cells, accompanied by an enhanced cell motility, and by the downregulation of E‑cadherin and the upregulation of Snail expression. By contrast, the ectopic overexpression of MBNL1 suppressed EMT, characterized by the upregulation of E‑cadherin and the downregulation of Snail expression. Mechanistically, Snail rather than E‑cadherin, was identified as a direct downstream target gene of MBNL1. The ectopic the overexpression of MBNL1 markedly enhanced the recruitment of Snail transcripts to processing bodies (P‑bodies), leading to the increased degradation of Snail mRNA and consequent translational silencing. Furthermore, the effect of MBNL1 on CRC cell migration was confirmed in additional CRC cell lines. SW480 and HT‑29 cells exhibited similar changes in migratory capacity and the expression of Snail/E‑cadherin to those observed in HCT‑116 cells. On the whole, this study demonstrates that MBNL1 destabilizes Snail transcripts and, in turn, suppresses the EMT of CRC cells through the Snail/E‑cadherin axis in vitro. Therefore, this EMT‑related MBNL1/Snail/E‑cadherin axis may prove to be a novel therapeutic target for CRC metastasis.

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1	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
2	Scheer A and Auer RA: Surveillance after curative resection of colorectal cancer. Clin Colon Rectal Surg. 22:242–250. 2009. View Article : Google Scholar :
3	Van Cutsem E, Nordlinger B and Cervantes A; ESMO Guidelines Working Group: Advanced colorectal cancer: ESMO Clinical Practice Guidelines for treatment. Ann Oncol. 21(Suppl 5): v93–v97. 2010. View Article : Google Scholar : PubMed/NCBI
4	Schüle S, Dittmar Y, Knösel T, Krieg P, Albrecht R, Settmacher U and Altendorf-Hofmann A: Long-term results and prognostic factors after resection of hepatic and pulmonary metastases of colorectal cancer. Int J Colorectal Dis. 28:537–545. 2013. View Article : Google Scholar
5	Brenner H, Kloor M and Pox CP: Colorectal cancer. Lancet. 383:1490–1502. 2014. View Article : Google Scholar
6	Fish L, Pencheva N, Goodarzi H, Tran H, Yoshida M and Tavazoie SF: Muscleblind-like 1 suppresses breast cancer metastatic colonization and stabilizes metastasis suppressor transcripts. Genes Dev. 30:386–398. 2016. View Article : Google Scholar : PubMed/NCBI
7	Cheng AW, Shi J, Wong P, Luo KL, Trepman P, Wang ET, Choi H, Burge CB and Lodish HF: Muscleblind-like 1 (Mbnl1) regulates pre-mRNA alternative splicing during terminal erythropoiesis. Blood. 124:598–610. 2014. View Article : Google Scholar : PubMed/NCBI
8	Vajda NA, Brimacombe KR, LeMasters KE and Ladd AN: Muscleblind-like 1 is a negative regulator of TGF-beta-dependent epithelial-mesenchymal transition of atrioventricular canal endocardial cells. Dev Dyn. 238:3266–3272. 2009. View Article : Google Scholar : PubMed/NCBI
9	Begemann G, Paricio N, Artero R, Kiss I, Pérez-Alonso M and Mlodzik M: muscleblind, a gene required for photoreceptor differentiation in Drosophila, encodes novel nuclear Cys3His-type zinc-finger-containing proteins. Development. 124:4321–4331. 1997.PubMed/NCBI
10	Artero R, Prokop A, Paricio N, Begemann G, Pueyo I, Mlodzik M, Perez-Alonso M and Baylies MK: The muscleblind gene participates in the organization of Z-bands and epidermal attachments of Drosophila muscles and is regulated by Dmef2. Dev Biol. 195:131–143. 1998. View Article : Google Scholar : PubMed/NCBI
11	Pascual M, Vicente M, Monferrer L and Artero R: The Muscleblind family of proteins: An emerging class of regulators of developmentally programmed alternative splicing. Differentiation. 74:65–80. 2006. View Article : Google Scholar : PubMed/NCBI
12	Wang ET, Cody NA, Jog S, Biancolella M, Wang TT, Treacy DJ, Luo S, Schroth GP, Housman DE, Reddy S, et al: Transcriptome-wide regulation of pre-mRNA splicing and mRNA localization by muscleblind proteins. Cell. 150:710–724. 2012. View Article : Google Scholar : PubMed/NCBI
13	Masuda A, Andersen HS, Doktor TK, Okamoto T, Ito M, Andresen BS and Ohno K: CUGBP1 and MBNL1 preferentially bind to 3′ UTRs and facilitate mRNA decay. Sci Rep. 2:2092012. View Article : Google Scholar
14	Batra R, Charizanis K, Manchanda M, Mohan A, Li M, Finn DJ, Goodwin M, Zhang C, Sobczak K, Thornton CA, et al: Loss of MBNL leads to disruption of developmentally regulated alternative polyadenylation in RNA-mediated disease. Mol Cell. 56:311–322. 2014. View Article : Google Scholar : PubMed/NCBI
15	Rau F, Freyermuth F, Fugier C, Villemin JP, Fischer MC, Jost B, Dembele D, Gourdon G, Nicole A, Duboc D, et al: Misregulation of miR-1 processing is associated with heart defects in myotonic dystrophy. Nat Struct Mol Biol. 18:840–845. 2011. View Article : Google Scholar : PubMed/NCBI
16	Bates RC and Mercurio AM: The epithelial-mesenchymal transition (EMT) and colorectal cancer progression. Cancer Biol Ther. 4:365–370. 2005. View Article : Google Scholar : PubMed/NCBI
17	Zeisberg M and Neilson EG: Biomarkers for epithelial-mesenchymal transitions. J Clin Invest. 119:1429–1437. 2009. View Article : Google Scholar : PubMed/NCBI
18	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
19	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
20	Weaver S, Dube S, Mir A, Qin J, Sun G, Ramakrishnan R, Jones RC and Livak KJ: Taking qPCR to a higher level: Analysis of CNV reveals the power of high throughput qPCR to enhance quantitative resolution. Methods. 50:271–276. 2010. View Article : Google Scholar : PubMed/NCBI
21	Yan JK, Zhu J, Gong ZZ, Wen J, Xiao YT, Zhang T and Cai W: Supplementary choline attenuates olive oil lipid emulsion-induced enterocyte apoptosis through suppression of CELF1/AIF pathway. J Cell Mol Med. 22:1562–1573. 2018. View Article : Google Scholar
22	Feng J, Cen J, Li J, Zhao R, Zhu C, Wang Z, Xie J and Tang W: Histone deacetylase inhibitor valproic acid (VPA) promotes the epithelial mesenchymal transition of colorectal cancer cells via up regulation of Snail. Cell Adhes Migr. 9:495–501. 2015. View Article : Google Scholar
23	Yang S, Li WS, Dong F, Sun HM, Wu B, Tan J, Zou WJ and Zhou DS: KITLG is a novel target of miR-34c that is associated with the inhibition of growth and invasion in colorectal cancer cells. J Cell Mol Med. 18:2092–2102. 2014. View Article : Google Scholar : PubMed/NCBI
24	Gupta GP and Massagué J: Cancer metastasis: Building a framework. Cell. 127:679–695. 2006. View Article : Google Scholar : PubMed/NCBI
25	Calon A, Espinet E, Palomo-Ponce S, Tauriello DV, Iglesias M, Céspedes MV, Sevillano M, Nadal C, Jung P, Zhang XH, et al: Dependency of colorectal cancer on a TGF-β-driven program in stromal cells for metastasis initiation. Cancer Cell. 22:571–584. 2012. View Article : Google Scholar : PubMed/NCBI
26	Valastyan S and Weinberg RA: Tumor metastasis: Molecular insights and evolving paradigms. Cell. 147:275–292. 2011. View Article : Google Scholar : PubMed/NCBI
27	Lin CH, Chiang MC and Chen YJ: MicroRNA-328 inhibits migration and epithelial-mesenchymal transition by targeting CD44 in nasopharyngeal carcinoma cells. OncoTargets Ther. 11:2375–2385. 2018. View Article : Google Scholar
28	Luo C, Yin D, Zhan H, Borjigin U, Li C, Zhou Z, Hu Z, Wang P, Sun Q, Fan J, et al: microRNA-501-3p suppresses metastasis and progression of hepatocellular carcinoma through targeting LIN7A. Cell Death Dis. 9:5352018. View Article : Google Scholar : PubMed/NCBI
29	Liu W, Li M, Chen X, Zhu S, Shi H, Zhang D, Cheng C and Li B: MicroRNA-1 suppresses proliferation, migration and invasion by targeting Notch2 in esophageal squamous cell carcinoma. Sci Rep. 8:51832018. View Article : Google Scholar : PubMed/NCBI
30	Zhang N, Tian L, Miao Z and Guo N: MicroRNA-197 induces epithelial-mesenchymal transition and invasion through the downregulation of HIPK2 in lung adenocarcinoma. J Genet. 47:47–53. 2018. View Article : Google Scholar : PubMed/NCBI
31	Brook JD, McCurrach ME, Harley HG, Buckler AJ, Church D, Aburatani H, Hunter K, Stanton VP, Thirion JP, Hudson T, et al: Molecular basis of myotonic dystrophy: Expansion of a trinucleotide (CTG) repeat at the 3′ end of a transcript encoding a protein kinase family member. Cell. 69:3851992.
32	Liquori CL, Ricker K, Moseley ML, Jacobsen JF, Kress W, Naylor SL, Day JW and Ranum LP: Myotonic dystrophy type 2 caused by a CCTG expansion in intron 1 of ZNF9. Science. 293:864–867. 2001. View Article : Google Scholar : PubMed/NCBI
33	Andres SF, Simmons JG, Mah AT, Santoro MA, Van Landeghem L and Lund PK: Insulin receptor isoform switching in intestinal stem cells, progenitors, differentiated lineages and tumors: Evidence that IR-B limits proliferation. J Cell Sci. 126:5645–5656. 2013. View Article : Google Scholar : PubMed/NCBI
34	The Human Protein Atlas: Expression of MBNL1 in cancer. https://www.proteinatlas.org/ENSG00000152601-MBNL1/pathology.
35	Paul S, Dansithong W, Jog SP, Holt I, Mittal S, Brook JD, Morris GE, Comai L and Reddy S: Expanded CUG repeats Dysregulate RNA splicing by altering the stoichiometry of the muscleblind 1 complex. J Biol Chem. 286:38427–38438. 2011. View Article : Google Scholar : PubMed/NCBI
36	Kulkarni M, Ozgur S and Stoecklin G: On track with P-bodies. Biochem Soc Trans. 38:242–251. 2010. View Article : Google Scholar : PubMed/NCBI
37	Chan SP and Slack FJ: microRNA-mediated silencing inside P-bodies. RNA Biol. 3:97–100. 2006. View Article : Google Scholar : PubMed/NCBI
38	Fillman C and Lykke-Andersen J: RNA decapping inside and outside of processing bodies. Curr Opin Cell Biol. 17:326–331. 2005. View Article : Google Scholar : PubMed/NCBI
39	Youn JY, Dunham WH, Hong SJ, Knight JDR, Bashkurov M, Chen GI, Bagci H, Rathod B, MacLeod G, Eng SWM, et al: High-Density Proximity Mapping Reveals the Subcellular Organization of mRNA-Associated Granules and Bodies. Mol Cell. 69:517–532, e11. 2018. View Article : Google Scholar : PubMed/NCBI
40	Hubstenberger A, Courel M, Bénard M, Souquere S, Ernoult-Lange M, Chouaib R, Yi Z, Morlot JB, Munier A, Fradet M, et al: P-Body Purification Reveals the Condensation of Repressed mRNA Regulons. Mol Cell. 68:144–157, e5. 2017. View Article : Google Scholar : PubMed/NCBI