بررسی جهش در اگزون7 ژن اکسین2 در سرطان کولورکتال (مطالعات آزمایشگاهی و بیوانفورماتیکی)

Background & Aims: Colorectal cancer is the third most common cancer in Iran. Wnt pathway changes are commonly reported in this type of cancer. Some of the important activities of this pathway are the role of carcinogenicity, cellular proliferation, cell migration, and so on. Axin2 acts as a negative regulator in the Wnt / TCF signaling pathway and helps in the formation of the beta-catenin destruction complex. Research has shown that this protein is mutated in many cases of CRC cancer.  In past studies, the mutation on the exon 7 of this gene is of particular importance because it is linked to PP2A phosphatase and in various scientific sources; mutations in this region have been reported repeatedly in colorectal tumors. The aim of this project is to study SNP with ID rs370618491 nucleotide C2140T in exon 7 of axin 2 gene and to investigate the possible role of mutation in this SNP in colorectal cancer. Materials & Methods: In this project, 147 frozen samples of patients (92 tumor tissues / 55 blood samples) and 25 control samples, and 3 cell lines of colorectal cancer, HT29, SW480 and CACO-2 were used. SNP selection was performed using bioinformatics methods including the use of NCBI, Ensembl databases and polyphen tool; DNA extraction was done by phenol-chloroform method. The PCR-RFLP method was used. The PCR-RFLP method was used and statistical analysis of demographic and clinical data was done using SPSS software. Results: This study showed that in one of 147 patient samples, there was a mutation in our preferred SNP position; in other samples, including cell lines no mutation showed. The results of SPSS 16.0 showed that the incidence of colorectal cancer in men was 55.7% and the mean age of patients over 40 years was 74%. It was also found that the size of the tumor (T) with P = 0.016 and the degree of tumor diffusion to the lymph nodes (N) with P=0.001 had a significant relationship with the grade of cancer progression. Conclusion: The presence one mutation in 147 samples indicates the low frequency of this SNP in the Iranian population. Therefore, despite the importance of exon 7 in axin 2, laboratory results were not consistent with the results of studies. Some factors can influence this outcome, including high variability in factors affecting colorectal cancer and high genetic variation in Iranian population. In addition, it usually comes from bioinformatics studies that provide information about the importance of an issue, not its prevalence in a particular population; therefore, bioinformatics studies alone are not enough and the frequency and clinical effects of a mutation in laboratory methods should also be considered. Due to the importance of axin 2 in the Wnt pathway and the high rates of mutations observed in previous studies, investigating the level of gene expression of axin 2 and the quality of its protein function can be important in future studies.