Optimization of High Resolution Melting (HRM) Method for Genotyping and Estimating the Frequency of SNP rs751402 on ERCC5 Gene in Vietnamese Breast Cancer Patients

Abstract

Breast cancer (BC) is the leading cancer-causing death for women worldwide. Among BC-causing genes, ERCC5 plays an important role in DNA damage repair. The cancer risk can be modified by Single Nucleotide Polymorphisms (SNPs), and thus, SNPs may be considered as potential markers of carcinogenesis. Among SNPs of the ERCC5 gene, rs751402 (A/G) had been found related to BC risk in the Chinese and Thailand populations. This study aims to develop the genotyping condition for rs751402 and then to predict the relation to BC risk in a Vietnamese population. 50 BC blood samples were examined. High Resolution Melting (HRM) genotyping method was optimized for rs751402. The allelic and genotypic frequencies of rs751402 in this study were compared with its data of Kinh Vietnamese in 1000Genomes as controls to predict the association with BC risk. The HRM genotyping assay was constructed with high sensitivity, specificity, and stability. The results revealed the minor allele frequency was 31.91%, indicating rs751402 is high polymorphism. The predicted association with BC showed rs751402 A allele tends to confer protection against BC (OR = 0.75) but not statistically significant (P-value = 0.28). Even so, rs75140 is still considered as a potential marker for further association studies with BC risk in the Vietnamese population due to its high polymorphism. 1000Genomes has certain reference value; however, some concern remains. Consequently, a further genotyping assay is proposed to conduct in a proper control group and larger sample size to confirm the relationship between rs751402 and BC risk in Vietnamese women.