Downregulation of miR-3127-5p promotes epithelial-mesenchymal transition via FZD4 regulation of Wnt/β-catenin signaling in non-small-cell lung cancer
Yang Yang
Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, Shanghai, China
Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, China
Search for more papers by this authorYifeng Sun
Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, China
Search for more papers by this authorYun Wu
Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, Shanghai, China
Search for more papers by this authorDongfang Tang
Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, Shanghai, China
Search for more papers by this authorXi Ding
Central Laboratory, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorWen Xu
Central Laboratory, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorCorresponding Author
Bo Su
Central Laboratory, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China
Correspondence
Wen Gao, Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, No. 221, Yananxi Road, Shanghai, 200040, China.
Email: [email protected]
Bo Su, Central Laboratory, Shanghai Pulmonary Hospital, Tongji University School of Medicine, No. 507, Zhengmin Road, Shanghai, 200433, China.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Wen Gao
Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, Shanghai, China
Correspondence
Wen Gao, Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, No. 221, Yananxi Road, Shanghai, 200040, China.
Email: [email protected]
Bo Su, Central Laboratory, Shanghai Pulmonary Hospital, Tongji University School of Medicine, No. 507, Zhengmin Road, Shanghai, 200433, China.
Email: [email protected]
Search for more papers by this authorYang Yang
Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, Shanghai, China
Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, China
Search for more papers by this authorYifeng Sun
Department of Thoracic Surgery, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, China
Search for more papers by this authorYun Wu
Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, Shanghai, China
Search for more papers by this authorDongfang Tang
Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, Shanghai, China
Search for more papers by this authorXi Ding
Central Laboratory, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorWen Xu
Central Laboratory, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China
Search for more papers by this authorCorresponding Author
Bo Su
Central Laboratory, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China
Correspondence
Wen Gao, Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, No. 221, Yananxi Road, Shanghai, 200040, China.
Email: [email protected]
Bo Su, Central Laboratory, Shanghai Pulmonary Hospital, Tongji University School of Medicine, No. 507, Zhengmin Road, Shanghai, 200433, China.
Email: [email protected]
Search for more papers by this authorCorresponding Author
Wen Gao
Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, Shanghai, China
Correspondence
Wen Gao, Department of Thoracic Surgery, Huadong Hospital Affiliated to FuDan University, No. 221, Yananxi Road, Shanghai, 200040, China.
Email: [email protected]
Bo Su, Central Laboratory, Shanghai Pulmonary Hospital, Tongji University School of Medicine, No. 507, Zhengmin Road, Shanghai, 200433, China.
Email: [email protected]
Search for more papers by this authorAbstract
MiR-3127-5p has been implicated as a tumor-suppressive microRNA (miRNA) in non-small-cell lung cancer (NSCLC) and its expression was associated with tumor recurrence and poor prognosis. The aim of this study was to determine whether miR-3127-5p regulates epithelial-mesenchymal transition (EMT) in NSCLC, and to investigate the underlying mechanisms. Using qRT-PCR, we examined the expression levels of miR-3127-5p in a cohort of primary NSCLC specimens with and without distant metastasis. We further performed a series of in vitro and in vivo experiments to investigate the effects and underlying mechanism of miR-3127-5p on EMT, cell migration, invasion, and adhesion in NSCLC. We found that metastatic NSCLC tissues showed markedly downregulated miR-3127-5p expression. Transforming growth factor-β1 (TGF-β1) treatment induced EMT in A549 and H1299 cells, and downregulation of miR-3127-5p could result in the similar effect. Mechanically, we demonstrated that frizzled-4 (FZD4) is a target gene and miR-3127-5p exerts its effects by regulating the Wnt/β-catenin signaling. In addition, the expression levels of FZD4 and miR-3127-5p were also negatively associated in both clinical and xenografted tumors. Overall, these findings suggest that downregulation of miR-3127-5p promotes EMT through activating the Wnt/FZD4/β-catenin signaling pathway and may represent a therapeutic target for NSCLC metastasis.
CONFLICTS OF INTEREST
The authors declare that there is no conflict of interest in this work.
Supporting Information
Additional Supporting Information may be found online in the supporting information tab for this article.
| Filename | Description |
|---|---|
| mc22805-sup-0001-SuppFig-S1.tif15 MB |
Figure S1. S1 Downregulation of miR-3127-5p promotes EMT in NSCLC cells. |
| mc22805-sup-0002-SuppTable-S1.docx13.4 KB |
Table S1. The primers used in qRT-PCR |
| mc22805-sup-0003-SuppLegend-S1.docx13.8 KB | Supporting Legend S1. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1 Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin. 2016; 66: 7–30.
- 2 Cufer T, Ovcaricek T, O'Brien ME. Systemic therapy of advanced non-small cell lung cancer: major-developments of the last 5-years. Eur J Cancer. 2013; 49: 1216–1225.
- 3 Olaussen KA, Postel-Vinay S. Predictors of chemotherapy efficacy in non-small-cell lung cancer: a challenging landscape. Ann Oncol. 2016; 27: 2004–2016.
- 4 Mehlen P, Puisieux A. Metastasis: a question of life or death. Nat Rev Cancer. 2006; 6: 449–458.
- 5 Yang J, Weinberg RA. Epithelial-mesenchymal transition: at the crossroads of development and tumor metastasis. Dev Cell. 2008; 14: 818–829.
- 6 da Silva SD, Morand GB, Alobaid FA, et al. Epithelial-mesenchymal transition (EMT) markers have prognostic impact in multiple primary oral squamous cell carcinoma. Clin Exp Metastasis. 2015; 32: 55–63.
- 7 Zeisberg M, Neilson EG. Biomarkers for epithelial-mesenchymal transitions. J Clin Invest. 2009; 119: 1429–1437.
- 8 Thiery JP, Sleeman JP. Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol. 2006; 7: 131–142.
- 9 Moody SE, Perez D, Pan TC, et al. The transcriptional repressor Snail promotes mammary tumor recurrence. Cancer Cell. 2005; 8: 197–209.
- 10 Dhawan P, Singh AB, Deane NG, et al. Claudin-1 regulates cellular transformation and metastatic behavior in colon cancer. J Clin Invest. 2005; 115: 1765–1776.
- 11 Huber MA, Azoitei N, Baumann B, et al. NF-kappaB is essential for epithelial-mesenchymal transition and metastasis in a model of breast cancer progression. J Clin Invest. 2004; 114: 569–581.
- 12 Anastas JN, Moon RT. WNT signalling pathways as therapeutic targets in cancer. Nat Rev Cancer. 2013; 13: 11–26.
- 13 Gurney A, Axelrod F, Bond CJ, et al. Wnt pathway inhibition via the targeting of Frizzled receptors results in decreased growth and tumorigenicity of human tumors. Proc Natl Acad Sci USA. 2012; 109: 11717–11722.
- 14 Akiri G, Cherian MM, Vijayakumar S, Liu G, Bafico A, Aaronson SA. Wnt pathway aberrations including autocrine Wnt activation occur at high frequency in human non-small-cell lung carcinoma. Oncogene. 2009; 28: 2163–2172.
- 15 Woenckhaus M, Merk J, Stoehr R, et al. Prognostic value of FHIT, CTNNB1, and MUC1 expression in non-small cell lung cancer. Hum Pathol. 2008; 39: 126–136.
- 16 Togashi Y, Hayashi H, Terashima M, et al. Inhibition of beta-Catenin enhances the anticancer effect of irreversible EGFR-TKI in EGFR-mutated non-small-cell lung cancer with a T790M mutation. J Thorac Oncol. 2015; 10: 93–101.
- 17 Wang H, Guan X, Tu Y, et al. MicroRNA-29b attenuates non-small cell lung cancer metastasis by targeting matrix metalloproteinase 2 and PTEN. J Exp Clin Cancer Res. 2015; 34: 59.
- 18 Zhai L, Ma C, Li W, Yang S. Liu Z. miR-143 suppresses epithelial-mesenchymal transition and inhibits tumor growth of breast cancer through down-regulation of ERK5. Mol Carcinog. 2016; 55: 1990–2000.
- 19 Xu W, Ji J, Xu Y, et al. MicroRNA-191, by promoting the EMT and increasing CSC-like properties, is involved in neoplastic and metastatic properties of transformed human bronchial epithelial cells. Mol Carcinog. 2015; 54: E148–E161.
- 20 Chen W, Wang J, Liu S, et al. MicroRNA-361-3p suppresses tumor cell proliferation and metastasis by directly targeting SH2B1 in NSCLC. J Exp Clin Cancer Res. 2016; 35: 76.
- 21 Sun Y, Chen C, Zhang P, et al. Reduced miR-3127-5p expression promotes NSCLC proliferation/invasion and contributes to dasatinib sensitivity via the c-Abl/Ras/ERK pathway. Sci Rep. 2014; 4: 6527.
- 22 Xu J, Lamouille S, Derynck R. TGF-beta-induced epithelial to mesenchymal transition. Cell Res. 2009; 19: 156–172.
- 23 Abba ML, Patil N, Leupold JH, et al. MicroRNAs as novel targets and tools in cancer therapy. Cancer Lett. 2017; 387: 84–94.
- 24 Cheng CJ, Bahal R, Babar IA, et al. MicroRNA silencing for cancer therapy targeted to the tumour microenvironment. Nature. 2015; 518: 107–110.
- 25 Xiao D, He J. Epithelial mesenchymal transition and lung cancer. J Thorac Dis 2010; 2: 154–159.
- 26 Prudkin L, Liu DD, Ozburn NC, et al. Epithelial-to-mesenchymal transition in the development and progression of adenocarcinoma and squamous cell carcinoma of the lung. Mod Pathol. 2009; 22: 668–678.
- 27 Soltermann A, Tischler V, Arbogast S, et al. Prognostic significance of epithelial-mesenchymal and mesenchymal-epithelial transition protein expression in non-small cell lung cancer. Clin Cancer Res. 2008; 14: 7430–7437.
- 28 Dong P, Kaneuchi M, Watari H, Sudo S, Sakuragi N. MicroRNA-106b modulates epithelial-mesenchymal transition by targeting TWIST1 in invasive endometrial cancer cell lines. Mol Carcinog. 2014; 53: 349–359.
- 29 Stewart DJ. Wnt signaling pathway in non-small cell lung cancer. J Natl Cancer Inst. 2014; 106.
- 30 Xu Z, Liu D, Fan C, Luan L, Zhang X, Wang E. DIXDC1 increases the invasion and migration ability of non-small-cell lung cancer cells via the PI3K-AKT/AP-1 pathway. Mol Carcinog. 2014; 53: 917–925.
- 31 Chen Y, Nowak I, Huang J, et al. Erk/MAP kinase signaling pathway and neuroendocrine differentiation of non-small-cell lung cancer. J Thorac Oncol. 2014; 9: 50–58.
- 32 Jeon SH, Yoon JY, Park YN, et al. Axin inhibits extracellular signal-regulated kinase pathway by Ras degradation via beta-catenin. J Biol Chem. 2007; 282: 14482–14492.
- 33 Jeong WJ, Yoon J, Park JC, et al. Ras stabilization through aberrant activation of Wnt/β-catenin signaling promotes intestinal tumorigenesis. Sci Signal. 2012; 5: ra30.
- 34 Labbé El, Letamendia Attisano AL. Association of Smads with lymphoid enhancer binding factor 1/T cell-specific factor mediates cooperative signaling by the transforming growth factor-beta and wnt pathways. Proc Natl Acad Sci USA. 2000; 97: 8358–8363.
- 35 Tickenbrock L, Hehn S, Sargin B, et al. Activation of Wnt signalling in acute myeloid leukemia by induction of Frizzled-4. Int J Oncol. 2008; 33: 1215–1221.
- 36 Pu X, Roth JA, Hildebrandt MA, et al. MicroRNA-related genetic variants associated with clinical outcomes in early-stage non-small cell lung cancer patients. Cancer Res. 2013; 73: 1867–1875.
- 37 Lin J, Zandi R, Shao R, et al. A miR-SNP biomarker linked to an increased lung cancer survival by miRNA-mediated down-regulation of FZD4 expression and Wnt signaling. Sci Rep. 2017; 7: 9029.
- 38 Kalluri R, Weinberg RA. The basics of epithelial-mesenchymal transition. J Clin Invest. 2009; 119: 1420–1428.
- 39 Xu Q, Wang Y, Dabdoub A, et al. Vascular development in the retina and inner ear: control by Norrin and Frizzled-4, a high-affinity ligand-receptor pair. Cell. 2004; 116: 883–895.
- 40 Reya T, Clevers H. Wnt signalling in stem cells and cancer. Nature. 2005; 434: 843–850.
- 41 Neth P, Ries C, Karow M, Egea V, Ilmer M, Jochum M. The Wnt signal transduction pathway in stem cells and cancer cells: influence on cellular invasion. Stem Cell Rev. 2007; 3: 18–29.
- 42 Gupta S, Iljin K, Sara H, et al. FZD4 as a mediator of ERG oncogene-induced WNT signaling and epithelial-to-mesenchymal transition in human prostate cancer cells. Cancer Res. 2010; 70: 6735–6745.
