Hsa-miR-424 by targeting APC increases Wnt signaling activates in colorectal cancer

Fasihi, Ali; Nemati, Hossein; Kabiri, Farnoush; Hasheminasab, Hoda; Mohamad Soltani, Bahram

Hsa-miR-424 by targeting APC increases Wnt signaling activates in colorectal cancer

Document Type : Original Research

Authors

Ali Fasihi ¹

Hossein Nemati ¹

Farnoush Kabiri ¹

Hoda Hasheminasab ¹

Bahram Mohamad Soltani ²

¹ Molecular Genetics Department, Faculty of Biological Sciences, Tarbiat Modares University

² Genetics Department Faculty of Biological Sciences

Abstract

The activity of Wnt signaling pathway is increased in colorectal cancer. For this reason, finding new positive and negative regulators for this pathway is a treatment and diagnostic strategy of colorectal cancer. Our bioinformatics analysis indicated that hsa-miR-424 (miR-424) could be a possible regulator of the Wnt signaling pathway. Accordingly, the expression level of miR-424 in colorectal cancer tissues was elevated compared with normal pairs and the results of RT-qPCR showed a significant increase in miR-424 expression (p < 0.01). Then, molecular analyzes using Top/Fop Flash and RT-qPCR techniques indicated that miR-424 overexpression leads to increased Wnt pathway activity in the SW480 cell line. In addition, the small molecules IWP-2 and PNU-74654 were used to inhibit the Wnt signaling pathway, and the miR-424 overexpression suggested that exert its effect on the level of β-catenin complex degradation. Then, dual-luciferase assay validated the interaction between miR-424 and APC. Overall, our results suggest miR-424 is a positive regulator of the Wnt signaling pathway, and it could be a possible prognosis for colorectal cancer.

Keywords

Wnt signaling pathway

Colorectal cancer

miR-424

Dual-luciferase assay

Subjects

Molecular biotechnology

1. De Klerk, C., et al., Socioeconomic and ethnic inequities within organised colorectal cancer screening programmes worldwide. Gut, 2018. 67(4): p. 679-687.
2. Arnold, M., et al., Global patterns and trends in colorectal cancer incidence and mortality. Gut, 2017. 66(4): p. 683-691.
3. Meyerhardt, J.A. and R.J. Mayer, Systemic therapy for colorectal cancer. New England journal of medicine, 2005. 352(5): p. 476-487.
4. Jiang, M.-C., et al., Emerging roles of lncRNA in cancer and therapeutic opportunities. American journal of cancer research, 2019. 9(7): p. 1354.
5. Hombach, S. and M. Kretz, Non-coding RNAs: classification, biology and functioning. Non-coding RNAs in colorectal cancer, 2016: p. 3-17.
6. Cai, Y., et al., A brief review on the mechanisms of miRNA regulation. Genomics, proteomics & bioinformatics, 2009. 7(4): p. 147-154.
7. Schatoff, E.M., B.I. Leach, and L.E. Dow, Wnt signaling and colorectal cancer. Current colorectal cancer reports, 2017. 13(2): p. 101-110.
8. Jung, Y.-S. and J.-I. Park, Wnt signaling in cancer: therapeutic targeting of Wnt signaling beyond β-catenin and the destruction complex. Experimental & Molecular Medicine, 2020. 52(2): p. 183-191.
9. Chen, H.-Y., et al., miR-103/107 prolong Wnt/β-catenin signaling and colorectal cancer stemness by targeting Axin2. Scientific reports, 2019. 9(1): p. 1-13.
10. Dai, W., et al., miR-424-5p promotes the proliferation and metastasis of colorectal cancer by directly targeting SCN4B. Pathology-Research and Practice, 2020. 216(1): p. 152731.
11. Li, N., CircTBL1XR1/miR-424 axis regulates Smad7 to promote the proliferation and metastasis of colorectal cancer. Journal of Gastrointestinal Oncology, 2020. 11(5): p. 918.
12. Cheng, C., et al., FENDRR sponges miR-424-5p to inhibit cell proliferation, migration and invasion in colorectal cancer. Technology in cancer research & treatment, 2020. 19: p. 1533033820980102.
13. Anastas, J.N. and R.T. Moon, WNT signalling pathways as therapeutic targets in cancer. Nature Reviews Cancer, 2013. 13(1): p. 11-26.
14. Clevers, H. and R. Nusse, Wnt/β-catenin signaling and disease. Cell, 2012. 149(6): p. 1192-1205.
15. Mohammadi, A., B. Mansoori, and B. Baradaran, The role of microRNAs in colorectal cancer. Biomedicine & Pharmacotherapy, 2016. 84: p. 705-713.
16. Najafi, H., et al., Alternative splicing of the OCC-1 gene generates three splice variants and a novel exonic microRNA, which regulate the Wnt signaling pathway. RNA, 2017. 23(1): p. 70-85.
17. Xu, X.M., et al., Expression of miR-21, miR-31, miR-96 and miR-135b is correlated with the clinical parameters of colorectal cancer. Oncology letters, 2012. 4(2): p. 339-345.
18. Hsieh, I.-S., et al., MicroRNA-320 suppresses the stem cell-like characteristics of prostate cancer cells by downregulating the Wnt/beta-catenin signaling pathway. Carcinogenesis, 2013. 34(3): p. 530-538.
19. Kim, N.H., et al., p53 and microRNA-34 are suppressors of canonical Wnt signaling. Science signaling, 2011. 4(197): p. ra71-ra71.
20. Kim, N.H., et al., p53 regulates nuclear GSK-3 levels through miR-34-mediated Axin2 suppression in colorectal cancer cells. Cell cycle, 2013. 12(10): p. 1578-1587.
21. Su, J., et al., MicroRNA-200a suppresses the Wnt/β-catenin signaling pathway by interacting with β-catenin. International journal of oncology, 2012. 40(4): p. 1162-1170.
22. Lewis, B.P., C.B. Burge, and D.P. Bartel, Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. cell, 2005. 120(1): p. 15-20.
23. Wei, S., et al., miR-424-5p promotes proliferation of gastric cancer by targeting Smad3 through TGF-β signaling pathway. Oncotarget, 2016. 7(46): p. 75185.
24. Xu, J., et al., CUL2 overexpression driven by CUL2/E2F1/miR-424 regulatory loop promotes HPV16 E7 induced cervical carcinogenesis. Oncotarget, 2016. 7(21): p. 31520.
25. Zhang, D., et al., Hypoxia-induced miR-424 decreases tumor sensitivity to chemotherapy by inhibiting apoptosis. Cell death & disease, 2014. 5(6): p. e1301-e1301.
26. Zhang, M., et al., MiR-424 promotes non-small cell lung cancer progression and metastasis through regulating the tumor suppressor gene TNFAIP1. Cellular Physiology and Biochemistry, 2017. 42(1): p. 211-221.
27. Rogers, H., et al., An investigation of WNT pathway activation and association with survival in central nervous system primitive neuroectodermal tumours (CNS PNET). British journal of cancer, 2009. 100(8): p. 1292-1302.
28. Voronkov, A. and S. Krauss, Wnt/beta-catenin signaling and small molecule inhibitors. Current pharmaceutical design, 2013. 19(4): p. 634-664.