High-resolution melting (HRM) analysis for the detection of single nucleotide polymorphisms in microRNA target sites

Background: The function of microRNAs (miRNAs) depends on the binding of miRNAs to their target sequences in the 3'UTR of messenger RNAs (mRNAs), which enhances the degradation of mRNAs and consequently, represses their expression. Single nucleotide polymorphisms (SNPs) in the miRNA target sequences may affect or impair the binding of miRNAs. Studies have shown that SNPs in miRNA target sites (miR-TS-SNPs) have a great influence on diverse biological functions, including pharmacogenomics and disease susceptibilities in human.

Methods: High-resolution melting (HRM) analysis was applied for investigating the allele frequencies of 3 miR-TS-SNPs (PLA2G2A, IL-16, and NOD2) in acute leukemia. We also compared the genotypes of acute lymphoblastic leukemia patients at initial diagnosis and complete remission.

Results: HRM analysis revealed 3 genotypes (both homozygous and heterozygous) in the 3 miR-TS-SNPs. The allele frequencies of all 3 miR-TS-SNPs were similar in normal individuals and patients with acute myelogenous leukemia. Most patients with acute lymphoblastic leukemia had the same genotypes at initial diagnosis and complete remission.

Conclusions: Large scale scanning of case-control studies for miR-TS-SNPs may contribute to the investigation of their roles and pathogenesis mechanisms in human diseases. Our study showed that HRM analysis can be an efficient tool for studies of miR-TS-SNPs.