Zhu Chaoyong, Odeberg Jacob, Hamsten Anders, Eriksson Per
Atherosclerosis Research Unit, King Gustav V Research Institute, Department of Medicine, Karolinska Institute, Stockholm, Sweden.
Biochem Biophys Res Commun. 2006 Sep 29;348(3):1150-6. doi: 10.1016/j.bbrc.2006.07.174. Epub 2006 Aug 4.
A common matrix metalloproteinases-3 (MMP-3) -1612 5A/6A promoter polymorphism is associated with risk for cardiovascular disease, rheumatoid arthritis, and other diseases. Here we used the haplotype chromatin immunoprecipitation method to study allele-specific MMP-3 expression under in vivo conditions in heterozygous THP-1 cells. Pyrosequencing was used to analyse the ratio of 5A-allele to 6A-allele after chromatin immunoprecipitation using an antibody against phosphorylated active RNA polymerase II. There was no allele-specific difference in transcriptional activity during basal conditions, i.e., in unstimulated monocytic THP-1 cells. However, after stimulation of MMP-3 expression by monocyte differentiation or incubation with IL-1beta, the haplotype containing the 5A-allele was associated with higher transcriptional activity compared with the 6A-containing haplotype. Electromobility shift assay demonstrated increased binding of nuclear proteins to the 5A-allele after monocyte differentiation. In conclusion, the common MMP-3 5A/6A promoter polymorphism appears to be functional only during specific environmental conditions involving inflammation.
常见的基质金属蛋白酶-3（MMP-3）-1612 5A/6A启动子多态性与心血管疾病、类风湿性关节炎及其他疾病的风险相关。在此，我们使用单倍型染色质免疫沉淀法研究杂合THP-1细胞在体内条件下的等位基因特异性MMP-3表达。使用针对磷酸化活性RNA聚合酶II的抗体进行染色质免疫沉淀后，采用焦磷酸测序法分析5A等位基因与6A等位基因的比例。在基础条件下，即在未刺激的单核细胞THP-1细胞中，转录活性不存在等位基因特异性差异。然而，在通过单核细胞分化或与IL-1β孵育刺激MMP-3表达后，与含6A的单倍型相比，含5A等位基因的单倍型与更高的转录活性相关。电泳迁移率变动分析表明，单核细胞分化后核蛋白与5A等位基因的结合增加。总之，常见的MMP-3 5A/6A启动子多态性似乎仅在涉及炎症的特定环境条件下才具有功能。
