Epithelial–mesenchymal transition (EMT) is the most common cause of death in colorectal cancer (CRC). In this study, we investigated the functional roles of miRNA-17-5p in EMT of CRC cells.
In order to determine if miRNA-17-5p regulated EMT, the precursors and inhibitors of miR-17-5p were transduced into four CRC cells. To evaluate the regulatory mechanism, we performed argonaute 2 (Ago2) immunoprecipitation (IP) and luciferase assay. In addition, we used an intra-splenic injection mouse model of BALB/c nude mice to investigate the metastatic potential of miRNA-17-5p in vivo.
The miRNA-17-5p expression was lower in primary CRC tissues with metastasis than in primary CRC tissues without metastasis in our RNA sequencing data of patient tissue. Real-time quantitative PCR revealed that miRNA-17-5p was inversely correlated with that of vimentin in five CRC cell lines. Over-expression of miRNA-17-5p decreased vimentin expression and inhibited cell migration and invasion in both LoVo and HT29 cells. However, inhibition of miRNA-17-5p showed the opposite effect. Ago2 IP and luciferase assay revealed that miRNA-17-5p directly bound to the 3′UTR of VIM mRNA. Furthermore, miRNA-17-5p inhibited the metastasis of CRC into liver in vivo.
Our results demonstrated that miRNA-17-5p regulates vimentin expression, thereby regulating metastasis of CRC.
Subscribe to Journal
Get full journal access for 1 year
only $20.79 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Hubbard, J. M. & Grothey, A. Colorectal cancer in 2014: progress in defining first-line and maintenance therapies. Nat. Rev. Clin. Oncol. 12, 73–74 (2015).
Ferlay, J., Shin, H. R., Bray, F., Forman, D., Mathers, C. & Parkin, D. M. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int. J. Cancer 127, 2893–2917 (2010).
Lee, Y. S., Kim, S. Y., Song, S. J., Hong, H. K., Lee, Y., Oh, B. Y. et al. Crosstalk between CCL7 and CCR3 promotes metastasis of colon cancer cells via ERK-JNK signaling pathways. Oncotarget 7, 36842–36853 (2016).
Siegel, R., Desantis, C. & Jemal, A. Colorectal cancer statistics, 2014. CA Cancer J. Clin. 64, 104–117 (2014).
Oh, B. Y., Kim, S. Y., Lee, Y. S., Hong, H. K., Kim, T. W., Kim, S. H. et al. Twist1-induced epithelial-mesenchymal transition according to microsatellite instability status in colon cancer cells. Oncotarget 7, 57066–57076 (2016).
Tang, J., Li, Y., Wang, J., Wen, Z., Lai, M. & Zhang, H. Molecular mechanisms of microRNAs in regulating epithelial-mesenchymal transitions in human cancers. Cancer Lett. 371, 301–313 (2016).
Lujambio, A. & Lowe, S. W. The microcosmos of cancer. Nature 482, 347–355 (2012).
Sun, B., Gu, X., Chen, Z. & Xiang, J. MiR-610 inhibits cell proliferation and invasion in colorectal cancer by repressing hepatoma-derived growth factor. Am. J. Cancer Res. 5, 3635–3644 (2015).
Feiersinger, F., Nolte, E., Wach, S., Rau, T. T., Vassos, N., Geppert, C. et al. MiRNA-21 expression decreases from primary tumors to liver metastases in colorectal carcinoma. PLoS ONE 11, e0148580 (2016).
Park, S. M., Gaur, A. B., Lengyel, E. & Peter, M. E. The miR-200 family determines the epithelial phenotype of cancer cells by targeting the E-cadherin repressors ZEB1 and ZEB2. Genes Dev. 22, 894–907 (2008).
Ahmed, F. E., Jeffries, C. D., Vos, P. W., Flake, G., Nuovo, G. J., Sinar, D. R. et al. Diagnostic microRNA markers for screening sporadic human colon cancer and active ulcerative colitis in stool and tissue. Cancer Genomics Proteom. 6, 281–295 (2009).
Fan, C., Lin, Y., Mao, Y., Huang, Z., Liu, A. Y., Ma, H. et al. MicroRNA-543 suppresses colorectal cancer growth and metastasis by targeting KRAS, MTA1 and HMGA2. Oncotarget 7, 21825–21839 (2016).
Gao, Y., Luo, L. H., Li, S. & Yang, C. miR-17 inhibitor suppressed osteosarcoma tumor growth and metastasis via increasing PTEN expression. Biochem. Biophys. Res. Commun. 444, 230–234 (2014).
Kandalam, M. M., Beta, M., Maheswari, U. K., Swaminathan, S. & Krishnakumar, S. Oncogenic microRNA 17-92 cluster is regulated by epithelial cell adhesion molecule and could be a potential therapeutic target in retinoblastoma. Mol. Vis. 18, 2279–2287 (2012).
Jiang, Z., Yin, J., Fu, W., Mo, Y., Pan, Y., Dai, L. et al. MiRNA 17 family regulates cisplatin-resistant and metastasis by targeting TGFbetaR2 in NSCLC. PLoS ONE 9, e94639 (2014).
Fan, M., Sethuraman, A., Brown, M., Sun, W. & Pfeffer, L. M. Systematic analysis of metastasis-associated genes identifies miR-17-5p as a metastatic suppressor of basal-like breast cancer. Breast Cancer Res. Treat. 146, 487–502 (2014).
Xi, X. P., Zhuang, J., Teng, M. J., Xia, L. J., Yang, M. Y., Liu, Q. G. et al. MicroRNA-17 induces epithelial-mesenchymal transition consistent with the cancer stem cell phenotype by regulating CYP7B1 expression in colon cancer. Int. J. Mol. Med. 38, 499–506 (2016).
Jiang, H., Wang, P., Wang, Q., Wang, B., Mu, J., Zhuang, X. et al. Quantitatively controlling expression of miR-17~92 determines colon tumor progression in a mouse tumor model. Am. J. Pathol. 184, 1355–1368 (2014).
Shin, C. H., Ryu, S. & Kim, H. H. hnRNPK-regulated PTOV1-AS1 modulates heme oxygenase-1 expression via miR-1207-5p. BMB Rep. 50, 220–225 (2017).
Lee, W. Y. & Cho, Y. B. Comparison of colorectal cancer in differentially established liver metastasis models. Anticancer Res. 34, 3321–3328 (2014).
Cerami, E., Gao, J., Dogrusoz, U., Gross, B. E., Sumer, S. O., Aksoy, B. A. et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. Cancer Discov. 2, 401–404 (2012).
Gao, J., Aksoy, B. A., Dogrusoz, U., Dresdner, G., Gross, B., Sumer, S. O. et al. Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci. Signal. 6, pl1 (2013).
Rupaimoole, R. & Slack, F. J. MicroRNA therapeutics: towards a new era for the management of cancer and other diseases. Nat. Rev. Drug Discov. 16, 203–222 (2017).
Biggar, K. K. & Storey, K. B. Insight into post-transcriptional gene regulation: stress-responsive microRNAs and their role in the environmental stress survival of tolerant animals. J. Exp. Biol. 218, 1281–1289 (2015).
Martin, H. C., Wani, S., Steptoe, A. L., Krishnan, K., Nones, K., Nourbakhsh, E. et al. Imperfect centered miRNA binding sites are common and can mediate repression of target mRNAs. Genome Biol. 15, R51 (2014).
Bartel, D. P. MicroRNAs: target recognition and regulatory functions. Cell 136, 215–233 (2009).
Ellwanger, D. C., Buttner, F. A., Mewes, H. W. & Stumpflen, V. The sufficient minimal set of miRNA seed types. Bioinformatics 27, 1346–1350 (2011).
Mullany, L. E., Herrick, J. S., Wolff, R. K. & Slattery, M. L. MicroRNA seed region length impact on target messenger RNA expression and survival in colorectal cancer. PLoS ONE 11, e0154177 (2016).
Wang, T., Lin, F., Sun, X., Jiang, L., Mao, R., Zhou, S. et al. HOXB8 enhances the proliferation and metastasis of colorectal cancer cells by promoting EMT via STAT3 activation. Cancer Cell Int. 19, 3 (2019).
Yan, K., Xu, X., Wu, T., Li, J., Cao, G., Li, Y. et al. Knockdown of PYCR1 inhibits proliferation, drug resistance and EMT in colorectal cancer cells by regulating STAT3-Mediated p38 MAPK and NF-kappaB signalling pathway. Biochem. Biophys. Res. Commun. 520, 486–491 (2019).
Chen, J., Gong, C., Mao, H., Li, Z., Fang, Z., Chen, Q. et al. E2F1/SP3/STAT6 axis is required for IL-4-induced epithelial-mesenchymal transition of colorectal cancer cells. Int. J. Oncol. 53, 567–578 (2018).
Li, Y., Zhao, Z., Xu, C., Zhou, Z., Zhu, Z. & You, T. HMGA2 induces transcription factor Slug expression to promote epithelial-to-mesenchymal transition and contributes to colon cancer progression. Cancer Lett. 355, 130–140 (2014).
Mansoori, B., Mohammadi, A., Naghizadeh, S., Gjerstorff, M., Shanehbandi, D., Shirjang, S. et al. miR-330 suppresses EMT and induces apoptosis by downregulating HMGA2 in human colorectal cancer. J. Cell Physiol. 235, 920–931 (2020).
Hu, F., Min, J., Cao, X., Liu, L., Ge, Z., Hu, J. et al. MiR-363-3p inhibits the epithelial-to-mesenchymal transition and suppresses metastasis in colorectal cancer by targeting Sox4. Biochem. Biophys. Res. Commun. 474, 35–42 (2016).
Mou, T. Y., Zhang, R. R. & Wang, Y. N. MiRNA-212 acts as a tumor-suppressor in colorectal carcinoma through targeting SOX4. Eur. Rev. Med. Pharm. Sci. 23, 10751–10760 (2019).
Shen, X., Hu, X., Mao, J., Wu, Y., Liu, H., Shen, J. et al. The long noncoding RNA TUG1 is required for TGF-beta/TWIST1/EMT-mediated metastasis in colorectal cancer cells. Cell Death Dis. 11, 65 (2020).
Hu, X., Li, Y. Q., Li, Q. G., Ma, Y. L., Peng, J. J. & Cai, S. J. Osteoglycin (OGN) reverses epithelial to mesenchymal transition and invasiveness in colorectal cancer via EGFR/Akt pathway. J. Exp. Clin. Cancer Res. 37, 41 (2018).
Xing, Y., Jing, H., Zhang, Y., Suo, J. & Qian, M. MicroRNA-141-3p affected proliferation, chemosensitivity, migration and invasion of colorectal cancer cells by targeting EGFR. Int. J. Biochem. Cell Biol. 118, 105643 (2020).
The biospecimens for this study were provided by the Samsung Medical Center BioBank (20130014 and 20140001).
Ethics approval and consent to participate
Ethics approval for animal use was obtained from the Samsung Medical Center on Laboratory Animals Committee (approval number: 20180129002). We carried out animal experiments in accordance with the ARRIVE reporting guideline and Samsung Medical Center on Laboratory Animals Committee’s guideline.
The data supporting the finding of this study are available within the article and are available from the corresponding authors upon request.
The authors declare no competing interests.
This paper was supported by the Sungkyun Research Fund, Sungkyunkwan University, 2018.
Note This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution 4.0 International (CC BY 4.0).
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Kim, T.W., Lee, Y.S., Yun, N.H. et al. MicroRNA-17-5p regulates EMT by targeting vimentin in colorectal cancer. Br J Cancer 123, 1123–1130 (2020). https://doi.org/10.1038/s41416-020-0940-5