Characterization of a prolactin gene polymorphism and its associations with systemic lupus erythematosus
Characterization of a prolactin gene polymorphism and its associations with systemic lupus erythematosus
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DOI：
10.1002/1529-0131(200110)44:10
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发表时间：
2001-10-01
影响因子：
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通讯作者：
Davis, JRE
中科院分区：
文献类型：
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作者：
Stevens, A;Ray, D;Davis, JRE
Objective.高泌乳素血症与系统性红斑狼疮（SLE）有关，但其机制尚不清楚。催乳素在T淋巴细胞中表达，并在替代启动子区域的控制下。我们描述了该启动子-1149位点的G/T单核苷酸多态性（SNP），并评估了其在SLE患者中的患病率。电泳迁移率变动分析（EMSA）进行，以确定催乳素启动子中的DNA蛋白复合物的形成。将含有G/T启动子等位基因的报告基因构建体瞬时转染到Jurkat T细胞系中用于测定转录活性。体外用植物血凝素（PH Lambda）处理外周血淋巴细胞（PBL），测定催乳素信使RNA（mRNA）水平。EMSA表明GATA相关转录因子的结合被位置-1149的G/T SNP改变。Jurkat细胞中的瞬时转染研究表明，G等位基因始终产生较高的启动子活性。PHA体外处理PBL诱导GG（-1149）基因型患者的催乳素mRNA比TT-1149基因型患者的催乳素mRNA增加更大。SLE患者队列中的疾病相关性研究表明，与对照组相比，催乳素-1149 G等位基因的频率增加。我们发现了一个功能显著的多态性，改变催乳素启动子的活性和mRNA水平的淋巴细胞。免疫细胞产生的局部催乳素改变可能通过影响T细胞功能而促进疾病进展。
Objective. Hyperprolactinemia is associated with Systemic lupus erythematosus (SLE), but the mechanism is unknown. Prolactin is expressed in T lymphocytes and is under the control of an alternative promoter region. We characterized a G/T single-nucleotide polymorphism (SNP) at position -1149 of this promoter and assessed its prevalence in patients with SLE.Methods. Electrophoretic mobility shift assays (EMSAs) were performed to determine DNA protein complex formation in the prolactin promoter. Transient transfection of reporter gene constructs containing the G/T promoter alleles into the Jurkat T cell line were used to determine transcription activity. Peripheral blood lymphocytes (PBLs) were treated in vitro with phytohemagglutinin (PH Lambda) to determine levels of prolactin messenger RNA (mRNA).Results. EMSAs indicated that binding of a GATA-related transcription factor was altered by the G/T SNP at position -1149. Transient transfection studies in Jurkat cells showed that the G allele consistently produced higher promoter activity. PHA treatment of PBLs in vitro induced a greater increment of prolactin mRNA from patients with the GG(-1149) genotype than from those with the TT-1149 genotype. Disease association studies in a cohort of SLE patients demonstrated an increased frequency of the prolactin -1149 G allele compared with control subjects.Conclusion. We found a functionally significant polymorphism that alters prolactin promoter activity and mRNA levels in the lymphocytes. Altered local prolactin production by immune cells may contribute to disease progression by affecting T cell function.