Comparison Between the Results of In Vitro RFLP Technique and Bioinformatics Programs in the Determination Of Genotypes CYP17 Gene In Breast Cancer Patients, Maysan/Southern Iraq

Certain CYP 17 gene allelic variants implicated in steroidogenesis may increase the risk of breast cancer. All earlier investigations used restriction enzymes in the lab for practically searching for genetic polymorphism of the CYP17 gene utilizing the RFLP approach. It is important to note that the presented research is the first to assess the outcomes of genetic variations in the CYP17 gene both practically and using bioinformatics software. The purpose of the presented work was to diagnose genetic alterations in the CYP17 gene and to evaluate the results of laboratory work and the bioinformatics program. The research included 24 patients with breast cancer and 24 control samples. The number and size of CYP17 fragments were determined using sequencing of CYP17 in NEBcutter bioinformatics software and in vitro by electrophoresis in agarose gel to RFLP product. The genotypes of the amplified CYP 17  product were determined in vitro by the RFLP method and by bioinformatics using Geneious Prime. The results of NEBcutter revealed there are simple differences in the number and size of fragments of the CYP 17 gene compared with the in vitro RFLP technique. The results of the RFLP technique in the laboratory found a single genotype CT and no significant difference between the two groups, compared with the result of the Geneious Prime bioinformatics program found two genotypes CC and CT. According to the findings of sequencing, patients with breast cancer had two unique mutations in the CYP17 gene (T261C and A insertion) compared to the control group. In conclusion, it is possible to use a bioinformatics program to accurately identify mutations and genotypes.