Development of a T-ARMS-PCR Assay for Detecting Genetic Polymorphism in the Catalase (rs7943316) Gene in the Iraqi Population with Breast Cancer

Abstract

Abstract Numerous investigations have demonstrated that oxidative stress is markedly increased in breast cancer patients compared to their healthy counterparts. Catalase (CAT), a crucial antioxidant enzyme, plays a pivotal role in safeguarding cells against oxidative damage initiated by reactive oxygen species (ROS). The CAT (rs7943316) gene encodes catalase, and certain genetic variations in this gene have been observed to modify catalase activity and levels. Such changes can lead to an altered response to oxidative stress, potentially increasing the risk of breast cancer. In light of this, a novel tetra-primer amplification-refractory mutation system (T-ARMS)-PCR assay was developed to investigate the possible correlation between the CAT (rs7943316) gene polymorphism and the development of breast cancer in patients. This study introduces a simple primer design method called T-ARMS for identifying the single nucleotide polymorphism (SNP) in the CAT (rs7943316) gene. This method employs a one-step PCR, which is faster, more cost-effective, and more precise than existing techniques. Sanger sequencing was performed to validate the accuracy of our findings. The T-ARMS-PCR assay revealed a significant association between the A/T allele of the CAT (rs7943316) gene and breast cancer. Specifically, individuals with the TT genotype had a higher risk of developing breast cancer than those with the AA genotype. The T allele frequency was greater among breast cancer patients than in the control group, and genotype frequencies were consistent with the principles of the Hardy-Weinberg Equilibrium. This study is the first to showcase a rapid, cost-effective, and high-throughput method for detecting the SNP in the CAT (rs7943316) gene. This method has the potential to be employed in large-scale clinical trials.