Investigating the possible roles of mutations in axin1 and axin2 genes in colorectal cancer

Document Type : Original Research

Authors
A. Jalalifar 1
L. Safavi 2
S. Moradifard 3
M. Banoei 4
M. Khalilollahi 2
S. Mohammad Ganji 1
1 Department of Molecular Medicine, Medical Biotechnology Institute, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
2 Department of Biology, North Tehran Branch, Islamic Azad University, Tehran, Iran.
3 Hudson Institute of Medical Research, Monash University, Melbourne, VIC, Australia
4 Department of Molecular Medicine, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Abstract
Introduction: Colorectal cancer (CRC) is one of the leading cancers, following skin, breast, and stomach cancers. This study aimed to investigate the relationship between mutations in axin1 and axin2 in association with CRC.

Methods: Our study contains 147 fresh frozen samples from CRC patients, 25 normal samples, and 3 cell lines, including HT29, SW480, and CACO-2. The chosen SNPs from databases are placed in exon 5 of axin1, in exon 2 of axin1, and in exon 7 of axin2. By PCR-RFLP method, mutated samples were identified and sequenced.

Results: The results showed that mutations in the single-nucleotide polymorphism (SNP) in axin2 were observed in 1 out of 147 patient samples (0.68%). In the three sequences examined in axin2 (exon 7), mutations in SNP with rs79024445 at A2052C were observed. Statistical analysis of clinical and pathological data of patients showed a significant relationship between the tumor size factor and grade of cancer (P=0.016) as well as the degree of tumor diffusion to the lymph nodes factor with a grade of cancer (P=0.001).
Conclusion: The multi-factorial nature of cancer, the high genetic diversity of the Iranian population, and the limited statistical population could affect these outcomes. The observed mutations in each sample can also indicate the importance of personalized medicine in studying diseases

Keywords

Subjects

1. Słoka, J., M. Madej, and B. Strzalka-Mrozik, Molecular Mechanisms of the Antitumor Effects of Mesalazine and Its Preventive Potential in Colorectal Cancer. Molecules, 2023;28(13): 5081.
2. Elmahdi, R., et al., Shared environment and colorectal cancer: A Nordic pedigree registry‐based cohort study. International Journal of Cancer, 2022;151(8): 1261-1269.
3. Sung, H., et al., Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians, 2021;71(3): 209-249.
4. Xi, Y. and P. Xu, Global colorectal cancer burden in 2020 and projections to 2040. Translational Oncology, 2021;14(10): 101174.
5. Hossain, M.S., et al., Colorectal Cancer: A Review of Carcinogenesis, Global Epidemiology, Current Challenges, Risk Factors, Preventive and Treatment Strategies. Cancers, 2022;14(7): 1732.
6. Patel, S.G., et al., The rising tide of early-onset colorectal cancer: a comprehensive review of epidemiology, clinical features, biology, risk factors, prevention, and early detection. The Lancet Gastroenterology & Hepatology, 2022.
7. Liu, J., et al., Wnt/β-catenin signalling: function, biological mechanisms, and therapeutic opportunities. Signal Transduction and Targeted Therapy, 2022;7(1): 1-23.
8. Yu, F., et al., Wnt/β-catenin signaling in cancers and targeted therapies. Signal Transduction and Targeted Therapy, 2021;6(1): 1-24.
9. Kühl, S.J. and M. Kühl, On the role of Wnt/β-catenin signaling in stem cells. Biochimica et Biophysica Acta (BBA)-General Subjects, 2013;1830(2): 2297-2306.
10. Gavagan, M., et al., The Wnt pathway scaffold protein Axin promotes signaling specificity by suppressing competing kinase reactions. bioRxiv, 2019: 768242.
11. Qiao, Y., et al., Axis inhibition protein 1 (Axin1) deletion–induced hepatocarcinogenesis requires intact β‐catenin but not notch cascade in mice. Hepatology, 2019;70(6): 2003-2017.
12. Miete, C., et al., Gαi2-induced conductin/axin2 condensates inhibit Wnt/β-catenin signaling and suppress cancer growth. Nature communications, 2022;13(1): 1-16.
13. Salahshor, S. and J. Woodgett, The links between axin and carcinogenesis. Journal of clinical pathology, 2005;58(3): 225-236.
14. Ji, Y., et al., Therapeutic strategies targeting Wnt/β‑catenin signaling for colorectal cancer. International Journal of Molecular Medicine, 2022;49(1): 1-17.
15. Dong, X., et al., Genomic structure, chromosome mapping and expression analysis of the human AXIN2 gene. Cytogenetic and Genome Research, 2001;93(1-2): 26-28.
16. Aghabozorgi, A.S., et al., The genetic factors associated with Wnt signaling pathway in colorectal cancer. Life Sciences, 2020;256: 118006.
17. Otero, L., et al., Variations in AXIN2 predict risk and prognosis of colorectal cancer. BDJ open, 2019;5(1): 1-6.
18. Moradifard, S., Z. Minuchehr, and S.M. Ganji, An investigation on the c‐MYC, AXIN1, and COL11A1 gene expression in colorectal cancer. Biotechnology and Applied Biochemistry, 2022;69(4): 1576-1586.
19. Aghabozorgi, A.S., et al., Role of adenomatous polyposis coli (APC) gene mutations in the pathogenesis of colorectal cancer; current status and perspectives. Biochimie, 2019;157: 64-71.
20. Mazzoni, S.M., et al., An AXIN2 mutant allele associated with predisposition to colorectal neoplasia has context-dependent effects on AXIN2 protein function. Neoplasia, 2015;17(5): 463-472.
21. Rivera, B., et al., A novel AXIN2 germline variant associated with attenuated FAP without signs of oligondontia or ectodermal dysplasia. European Journal of Human Genetics, 2014;22(3): 423-426.
22. Wodarz, A. and R. Nusse, Mechanisms of Wnt signaling in development. Annual review of cell and developmental biology, 1998;14(1): 59-88.
23. Voronkov, A. and S. Krauss, Wnt/beta-catenin signaling and small molecule inhibitors. Current pharmaceutical design, 2013;19(4): 634-664.
24. Adzhubei, I.A., et al., A method and server for predicting damaging missense mutations. Nat Methods, 2010;7(4): 248-9.
25. Pettersen, E.F., et al., UCSF Chimera--a visualization system for exploratory research and analysis. J Comput Chem, 2004;25(13): 1605-12.
26. Guex, N. and M.C. Peitsch, SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis, 1997;18(15): 2714-23.
27. Vincze, T., J. Posfai, and R.J. Roberts, NEBcutter: a program to cleave DNA with restriction enzymes. Nucleic acids research, 2003;31(13): 3688-3691.
28. Rychlik, W., OLIGO 7 primer analysis software. Methods Mol Biol, 2007;402: 35-60.
29. Nong, J., et al., Phase separation of Axin organizes the β-catenin destruction complex. J Cell Biol, 2021;220(4).
30. Mallick, A., et al., Axin Family of scaffolding proteins in development: Lessons from C. elegans. Journal of developmental biology, 2019;7(4): 20.
31. Jin, L.H., et al., Detection of point mutations of the Axin1 gene in colorectal cancers. International journal of cancer, 2003;107(5): 696-699.
32. Khan, N.P., et al., Novelty of Axin 2 and lack of Axin 1 gene mutation in colorectal cancer: a study in Kashmiri population. Molecular and cellular biochemistry, 2011;355(1): 149-155.
33. Webster, M.T., et al., Sequence variants of the axin gene in breast, colon, and other cancers: an analysis of mutations that interfere with GSK3 binding. Genes, Chromosomes and Cancer, 2000;28(4): 443-453.
34. Sidore, C., et al., Genome sequencing elucidates Sardinian genetic architecture and augments association analyses for lipid and blood inflammatory markers. Nature genetics, 2015;47(11): 1272-1281.
35. Peterlongo, P., et al., Germline mutations of AXIN2 are not associated with nonsyndromic colorectal cancer. Human mutation, 2005;25(5): 498.
36. Dajani, R., et al., Structural basis for recruitment of glycogen synthase kinase 3β to the axin—APC scaffold complex. The EMBO journal, 2003;22(3): 494-501.
Pathobiology Reserach
Volume 28, Issue 2
March 2025
Pages 13-25