Analysis of Multiple Single Nucleotide Polymorphisms (SNPs) of Myeloperoxidase (MPO) to Screen for Genetic Markers Associated with Acute Leukemia in Chinese Han Population

Abstract

Myeloperoxidase (MPO) is an important metabolizing enzyme involved in oxidative stress responses to some environmental carcinogens including benzene, a chemical associated with bone marrow toxicity and leukemia. A novel multiplex polymerase chain reaction (PCR) technique based on adapter-ligation mediated allele-specific amplification (ALM-ASA) is simple, inexpensive and especially useful for multiplex single nucleotide polymorphisms (SNPs) genotyping. The purpose of this study was to examine the genetic markers associated with acute leukemia from multiple SNPs of MPO found in Chinese Han population. A case-control study was conducted, with 135 patients with adult acute leukemia and 187 cancer-free subjects as a control group. Nine SNPs in promoter and exons were selected that were markers related to gene function and risk of leukemia development. The multiplex PCR technique based on ALM-ASA was used to detect multiple SNP genotypes. The accuracy of ALM-ASA was tested by sequence and PCRrestriction fragment length polymorphism (RFLP) genotyping analysis. No polymorphisms were found at −1468T > C, −1053T > C, 958C > T, 967C > A, 1033 T > C. A marked linkage disequilibrium was found among −1816, −586, and −463 genotypes. Three haplotypes were constructed. SNP and haplotypes analysis showed the frequency of the MPO −463G > A mutant genotypes (G/A+A/A) was significantly less in cancer cases. The results of sequence and PCR-RFLP yielded results similar to ALM-ASA. Data suggest that MPO G-463A mutant genotypes may be associated with a reduced risk of acute leukemia development due to diminished activation of carcinogens. MPO G-463A variant may possess a protective genetic marker against acute leukemia in the Chinese Han population.

This study was supported by Jiangsu Province Medicine Foundation (H200538) and A Foundation of Graduate Creative Program of Jiang Su ([2005]7). Yue-Pu Pu contributed to the concept, design, and final approval of the article and obtained a funding source. Li-Hong Yin provided analysis and interpretation of data and administrative support. Fang-Yan Zhu collected and assembled data; Guo-Hua Zhou, Wei-Peng Wang, and Juan Luo designed the amplification primer and condition.