Shin Jeong-Ah, Chang Hun Soo, Park Se-Min, Jang An-Soo, Park Sung Woo, Park Jong Sook, Uh Soo-Taek, Il Lim Gune, Rhim Taiyoun, Kim Mi-Kyeong, Choi Inseon S, Chung Il Yup, Park Byung Lae, Shin Hyoung Doo, Park Choon-Sik
Genome Research Center for Allergy and Respiratory Diseases, Soonchunhyang University Bucheon Hospital, 1174, Jung Dong, Wonmi Ku, Bucheon, Gyeonggi Do, 420-767, Korea.
BMC Med Genet. 2009 Oct 20;10:106. doi: 10.1186/1471-2350-10-106.
We previously demonstrated that single nucleotide polymorphism (SNP) and haplotypes were associated with aspirin hypersensitivity in asthmatics. We investigated the genetic effects of the SNPs and haplotypes on the expression of the CysLTR2 gene.
We measured CysLTR2 protein and mRNA expression in EB virus-infected B cell lines from asthmatics having ht1+/+ and ht2+/+. A gel retardation assay was used to identify nuclear protein binding to the c.-819 promoter site. The function of promoter and 3'-UTR were assessed using pGL3 luciferase and pEGFP reporter system, respectively.
We found that the expression of CysLTR2 protein was higher in B cell lines of asthmatics having ht2+/+ than in those having ht1+/+. PMA/ionomycin induced higher mRNA expression of CysLTR2 in B cell lines from ht2+/+ asthmatics than those from ht1+/+ asthmatics. A nuclear protein from the B cell lines showed stronger DNA binding affinity with a probe containing c.-819T than one containing c.-819G. The luciferase activity of the c.-819T type of CysLTR2 promoter was higher than that of the c.-819G type. EGFP expression was higher in the EGFP-c.2078T 3'-UTR fusion construct than in the c.2078C construct.
The sequence variants of CysLTR2 may affect its transcription and the stability of its mRNA, resulting in altered expression of CysLTR2 protein, which in turn causes some asthmatics to be susceptible to aspirin hypersensitivity.
我们之前证明单核苷酸多态性（SNP）和单倍型与哮喘患者的阿司匹林超敏反应相关。我们研究了这些SNP和单倍型对CysLTR2基因表达的遗传效应。
我们测量了来自ht1+/+和ht2+/+哮喘患者的EB病毒感染B细胞系中CysLTR2蛋白和mRNA的表达。采用凝胶阻滞试验鉴定与c.-819启动子位点结合的核蛋白。分别使用pGL3荧光素酶和pEGFP报告系统评估启动子和3'-UTR的功能。
我们发现，ht2+/+哮喘患者的B细胞系中CysLTR2蛋白的表达高于ht1+/+患者。佛波酯/离子霉素诱导ht2+/+哮喘患者B细胞系中CysLTR2的mRNA表达高于ht1+/+哮喘患者。B细胞系的核蛋白与含有c.-819T的探针的DNA结合亲和力强于含有c.-819G的探针。CysLTR2启动子的c.-819T型荧光素酶活性高于c.-819G型。EGFP-c.2078T 3'-UTR融合构建体中的EGFP表达高于c.2078C构建体。
CysLTR2的序列变异可能影响其转录及其mRNA的稳定性，导致CysLTR2蛋白表达改变，进而使一些哮喘患者易患阿司匹林超敏反应。
