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Molecular Diagnostics

Clinical assessment of the miR-34, miR-200, ZEB1 and SNAIL EMT regulation hub underlines the differential prognostic value of EMT miRs to drive mesenchymal transition and prognosis in resected NSCLC



Patients with non-small cell lung cancer (NSCLC) receiving curative surgery have a risk of relapse, and adjuvant treatments only translate into a 5% increase in 5-year survival. We assessed the clinical significance of epithelial–mesenchymal transition (EMT) and explored its association with the [SNAIL/miR-34]:[ZEB/miR-200] regulation hub to refine prognostic information.


We validated a 7-gene EMT score using a consecutive series of 176 resected NSCLC. We quantified EMT transcription factors, microRNAs (miRs) of the miR-200, miR-34 families and miR-200 promoter hypermethylation to identify outcome predictors.


Most tumours presented with an EMT-hybrid state and the EMT score was not predictive of outcome. Individually, all miR-200 were inversely associated with the EMT score, but only chromosome-1 miRs, miR-200a, b, 429, were associated with disease-free survival (p = 0.08, 0.05 and 0.025) and overall survival (p = 0.013, 0.003 and 0.006). We validated these associations on The Cancer Genome Atlas data. Tumour unsupervised clustering based on miR expression identified two good prognostic groups, unrelated to the EMT score, suggesting that miR profiling may have an important clinical value.


miR-200 family members do not have similar predictive value. Core EMT-miR, regulators and not EMT itself, identify NSCLC patients with a low risk of relapse after surgery.

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Fig. 1: Correlation matrix of miR-34, miR-200, promoter methylation, epithelial and mesenchymal marker expression.
Fig. 2: Unsupervised hierarchical clustering based on miR-34, miR-200 expression and promoter methylation status.
Fig. 3: Kaplan–Meier OS and RFS plots.
Fig. 4: Kaplan–Meier OS and RFS plots.


  1. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65:87–108.

    Article  Google Scholar 

  2. Noone A, Howlader N, Krapcho M, Miller D, Brest A, Yu M, et al. (eds). SEER Cancer Statistics Review, 1975–2015. Bethesda:National cancer Institute; 2018.

  3. Wu Y-L, Tsuboi M, He J, John T, Grohe C, Majem M, et al. Osimertinib in resected EGFR-mutated non-small-cell lung cancer. N. Engl J Med. 2020;383:1711–23.

    Article  CAS  Google Scholar 

  4. Legras A, Pécuchet N, Imbeaud S, Pallier K, Didelot A, Roussel H, et al. Epithelial-to-mesenchymal transition and micrornas in lung cancer. Cancers. 2017;9:101.

  5. Kalluri R, Weinberg RA. The basics of epithelial-mesenchymal transition. J Clin Invest. 2009;119:1420–8.

    Article  CAS  Google Scholar 

  6. Thiery JP, Acloque H, Huang RYJ, Nieto MA. Epithelial-mesenchymal transitions in development and disease. Cell. 2009;139:871–90.

    Article  CAS  Google Scholar 

  7. Zaravinos A. The regulatory role of microRNAs in EMT and cancer. J Oncol. 2015;2015:865816.

    Article  Google Scholar 

  8. Zavadil J, Böttinger EP. TGF-beta and epithelial-to-mesenchymal transitions. Oncogene. 2005;24:5764–74.

    Article  CAS  Google Scholar 

  9. Lo H-W, Hsu S-C, Xia W, Cao X, Shih J-Y, Wei Y, et al. Epidermal growth factor receptor cooperates with signal transducer and activator of transcription 3 to induce epithelial-mesenchymal transition in cancer cells via up-regulation of TWIST gene expression. Cancer Res. 2007;67:9066–76.

    Article  CAS  Google Scholar 

  10. Tania M, Khan MA, Fu J. Epithelial to mesenchymal transition inducing transcription factors and metastatic cancer. Tumour Biol. 2014;35:7335–42.

    Article  CAS  Google Scholar 

  11. Zhang P, Sun Y, Ma L. ZEB1: at the crossroads of epithelial-mesenchymal transition, metastasis and therapy resistance. Cell Cycle. 2015;14:481–7.

    Article  CAS  Google Scholar 

  12. Gemmill RM, Roche J, Potiron VA, Nasarre P, Mitas M, Coldren CD, et al. ZEB1-responsive genes in non-small cell lung cancer. Cancer Lett. 2011;300:66–78.

    Article  CAS  Google Scholar 

  13. Pallier K, Cessot A, Côté J-F, Just P-A, Cazes A, Fabre E, et al. TWIST1 a new determinant of epithelial to mesenchymal transition in EGFR mutated lung adenocarcinoma. PLoS ONE. 2012;7:e29954.

    Article  CAS  Google Scholar 

  14. Gregory PA, Bracken CP, Bert AG, Goodall GJ. MicroRNAs as regulators of epithelial-mesenchymal transition. Cell Cycle. 2008;7:3112–8.

    Article  CAS  Google Scholar 

  15. Meng F, Wu G. The rejuvenated scenario of epithelial-mesenchymal transition (EMT) and cancer metastasis. Cancer Metastasis Rev. 2012;31:455–67.

    Article  CAS  Google Scholar 

  16. Pecot CV, Rupaimoole R, Yang D, Akbani R, Ivan C, Lu C, et al. Tumour angiogenesis regulation by the miR-200 family. Nat Commun. 2013;4:2427.

    Article  CAS  Google Scholar 

  17. Chen L, Gibbons DL, Goswami S, Cortez MA, Ahn Y-H, Byers LA, et al. Metastasis is regulated via microRNA-200/ZEB1 axis control of tumour cell PD-L1 expression and intratumoral immunosuppression. Nat Commun. 2014;5:5241.

    Article  CAS  Google Scholar 

  18. Lang Y, Xu S, Ma J, Wu J, Jin S, Cao S, et al. MicroRNA-429 induces tumorigenesis of human non-small cell lung cancer cells and targets multiple tumor suppressor genes. Biochem Biophys Res Commun. 2014;450:154–9.

    Article  CAS  Google Scholar 

  19. Title AC, Hong S-J, Pires ND, Hasenöhrl L, Godbersen S, Stokar-Regenscheit N, et al. Genetic dissection of the miR-200–Zeb1 axis reveals its importance in tumor differentiation and invasion. Nat Commun. 2018;9:4671.

    Article  CAS  Google Scholar 

  20. Damiano V, Brisotto G, Borgna S, di Gennaro A, Armellin M, Perin T, et al. Epigenetic silencing of miR-200c in breast cancer is associated with aggressiveness and is modulated by ZEB1: MIR-200C LOCUS METHYLATION IN BREAST CANCER. Genes Chromosomes Cancer. 2017;56:147–58.

    Article  CAS  Google Scholar 

  21. Pichler M, Ress AL, Winter E, Stiegelbauer V, Karbiener M, Schwarzenbacher D, et al. MiR-200a regulates epithelial to mesenchymal transition-related gene expression and determines prognosis in colorectal cancer patients. Br J Cancer. 2014;110:1614–21.

    Article  CAS  Google Scholar 

  22. Xue B, Chuang C-H, Prosser HM, Fuziwara CS, Chan C, Sahasrabudhe N, et al. miR-200 deficiency promotes lung cancer metastasis by activating cancer-associated fibroblasts. bioRxiv:2020.09.02.276550 [Preprint]. 2020.

  23. Tejero R, Navarro A, Campayo M, Viñolas N, Marrades RM, Cordeiro A, et al. miR-141 and miR-200c as markers of overall survival in early stage non-small cell lung cancer adenocarcinoma. PLoS ONE. 2014;9:e101899.

  24. On behalf of the EMT International Association (TEMTIA), Yang J, Antin P, Berx G, Blanpain C, Brabletz T, et al. Guidelines and definitions for research on epithelial–mesenchymal transition. Nat Rev Mol Cell Biol. 2020;21:341–52.

    Article  CAS  Google Scholar 

  25. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 2001;25(Dec):402–8.

    Article  CAS  Google Scholar 

  26. Byers LA, Diao L, Wang J, Saintigny P, Girard L, Peyton M, et al. An epithelial-mesenchymal transition gene signature predicts resistance to EGFR and PI3K inhibitors and identifies Axl as a therapeutic target for overcoming EGFR inhibitor resistance. Clin Cancer Res. 2013;19:279–90.

    Article  CAS  Google Scholar 

  27. Tan TZ, Miow QH, Miki Y, Noda T, Mori S, Huang RY-J, et al. Epithelial-mesenchymal transition spectrum quantification and its efficacy in deciphering survival and drug responses of cancer patients. EMBO Mol Med. 2014;6:1279–93.

    Article  CAS  Google Scholar 

  28. Chakraborty P, George JT, Tripathi S, Levine H, Jolly MK. Comparative study of transcriptomics-based scoring metrics for the epithelial-hybrid-mesenchymal spectrum. Front Bioeng Biotechnol. 2020;8:220.

  29. Pignon J-P, Tribodet H, Scagliotti GV, Douillard J-Y, Shepherd FA, Stephens RJ, et al. Lung adjuvant cisplatin evaluation: a pooled analysis by the LACE Collaborative Group. J Clin Oncol. 2008;26:3552–9.

    Article  Google Scholar 

  30. NSCLC Meta-analyses Collaborative Group, Arriagada R, Auperin A, Burdett S, Higgins JP, Johnson DH, et al. Adjuvant chemotherapy, with or without postoperative radiotherapy, in operable non-small-cell lung cancer: two meta-analyses of individual patient data. Lancet. 2010;375:1267–77.

    Article  CAS  Google Scholar 

  31. Jolly MK, Boareto M, Huang B, Jia D, Lu M, Ben-Jacob E, et al. Implications of the hybrid epithelial/mesenchymal phenotype in metastasis. Front Oncol. 2015;5:155.

    Article  Google Scholar 

  32. Young KC, Sangmin C, Taeyeong K, Jonathan A, Sarita A, Wade I, et al. Epithelial-mesenchymal transition (EMT) signature is inversely associated with T-cell infiltration in non-small cell lung cancer (NSCLC). Sci Rep. 2018;8:2918.

  33. Thompson JC, Hwang W-T, Davis C, Deshpande C, Jeffries S, Rajpurohit Y, et al. Gene signatures of tumor inflammation and epithelial-to-mesenchymal transition (EMT) predict responses to immune checkpoint blockade in lung cancer with high accuracy. Lung Cancer. 2020;139:1–8.

    Article  Google Scholar 

  34. Jolly MK, Boareto M, Huang B, Jia D, Lu M, Ben-Jacob E, et al. Implications of the hybrid epithelial/mesenchymal phenotype in metastasis. Front Oncol. 2015;5:155.

    Article  Google Scholar 

  35. Bracken CP, Gregory PA, Kolesnikoff N, Bert AG, Wang J, Shannon MF, et al. A double-negative feedback loop between ZEB1-SIP1 and the microRNA-200 family regulates epithelial-mesenchymal transition. Cancer Res. 2008;68:7846–54.

    Article  CAS  Google Scholar 

  36. Hill L, Browne G, Tulchinsky E. ZEB/miR-200 feedback loop: at the crossroads of signal transduction in cancer. Int J Cancer. 2013;132:745–54.

    Article  CAS  Google Scholar 

  37. Lindner P, Paul S, Eckstein M, Hampel C, Muenzner JK, Erlenbach-Wuensch K, et al. EMT transcription factor ZEB1 alters the epigenetic landscape of colorectal cancer cells. Cell Death Dis. 2020;11:147.

    Article  CAS  Google Scholar 

  38. Kim JS, Kim EJ, Lee S, Tan X, Liu X, Park S, et al. MiR-34a and miR-34b/c have distinct effects on the suppression of lung adenocarcinomas. Exp Mol Med. 2019;51:9.

  39. Kim T, Veronese A, Pichiorri F, Lee TJ, Jeon Y-J, Volinia S, et al. p53 regulates epithelial–mesenchymal transition through microRNAs targeting ZEB1 and ZEB2. J Exp Med. 2011;208:875–83.

    Article  CAS  Google Scholar 

  40. Xiao P, Liu W, Zhou H. miR-429 promotes the proliferation of non-small cell lung cancer cells via targeting DLC-1. Oncol Lett. 2016;12:2163–8.

    Article  CAS  Google Scholar 

  41. Mei Z, He Y, Feng J, Shi J, Du Y, Qian L, et al. MicroRNA-141 promotes the proliferation of non-small cell lung cancer cells by regulating expression of PHLPP1 and PHLPP2. FEBS Lett. 2014;588:3055–61.

    Article  CAS  Google Scholar 

  42. Zhu W, He J, Chen D, Zhang B, Xu L, Ma H, et al. Expression of miR-29c, miR-93, and miR-429 as potential biomarkers for detection of early stage non-small lung cancer. PLoS ONE. 2014;9:e87780.

    Article  CAS  Google Scholar 

  43. Wang F, Zhang L, Xu H, Li R, Xu L, Qin Z, et al. The significance role of microRNA-200c as a prognostic factor in various human solid malignant neoplasms: a meta-analysis. J Cancer. 2019;10:277–86.

    Article  CAS  Google Scholar 

  44. De Sousa E, Melo F, Wang X, Jansen M, Fessler E, Trinh A, et al. Poor-prognosis colon cancer is defined by a molecularly distinct subtype and develops from serrated precursor lesions. Nat Med. 2013;19:614–8.

    Article  CAS  Google Scholar 

  45. Noman MZ, Janji B, Abdou A, Hasmim M, Terry S, Tan TZ, et al. The immune checkpoint ligand PD-L1 is upregulated in EMT-activated human breast cancer cells by a mechanism involving ZEB-1 and miR-200. Oncoimmunology 2017;6:e1263412.

    Article  CAS  Google Scholar 

  46. Tulchinsky E, Demidov O, Kriajevska M, Barlev NA, Imyanitov E. EMT: a mechanism for escape from EGFR-targeted therapy in lung cancer. Biochim Biophys Acta. 2019;1871:29–39.

    CAS  Google Scholar 

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We thank Claudia DeToma, Marine Largeau, Elodie Michel and Lauriane Chambolle for their work and implication at the Biological Resources centre and Tumour Bank Platform PRB-HEGP (BB-0033-00063).


This study was supported by a grant from the integrated site of cancer research ‘cancer research for personalised medicine’ (SIRIC CARPEM). AL received personal research grants from Fondation de la Recherche Médicale. We thank the Tumour Bank Platform (PRB-HEGP BB-0033-00063) for its support.

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Conception or design: HB, SG and AL. Data collection: J-BL, FLP-B, AL, LG, CB, TD and AP. Data analysis: SG and AL. Redaction: AL, SG and HB. Final approval: SG, AD, TD, AP, GB, J-BL, J-BO, LG, CB, FLP-B, PL-P, AL and HB.

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Correspondence to Helene Blons.

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Ethics approval and consent to participate

This study, conducted at the European Georges Pompidou hospital, was approved by CPP Ile de France 2 ethics committee (nos. 2012-08-09 and 2012-08-09 A1) and registered in clinical (NCT03509779). Patients with NSCLC treated by surgery for curative intent signed informed consent for research and tumour tissues banking. Samples were stored frozen (−80 °C) at the Biological Resources centre and Tumour Bank Platform (PRB-HEGP BB-0033-00063).

Competing interests

AL reports grants from Fondation de la Recherche Médicale, during the conduct of the study. HB reports a grant from Site de la Recherche Intégrée sur le Cancer (SIRIC) CARPEM for this study.

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Garinet, S., Didelot, A., Denize, T. et al. Clinical assessment of the miR-34, miR-200, ZEB1 and SNAIL EMT regulation hub underlines the differential prognostic value of EMT miRs to drive mesenchymal transition and prognosis in resected NSCLC. Br J Cancer 125, 1544–1551 (2021).

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