Association of MMP-1 promoter polymorphism with risk of apical periodontitis.

转录因子Ets-1在MMP-1 mRNA表达中起重要作用。1VIMP-1基因启动子区-1607bp的单核苷酸多态性对Ets-1结合活性有影响。方法：将人牙周膜细胞接种于含10%FBS的α-MEM培养皿中，培养10天后，取细胞悬液，用MTT法检测Ets-1的结合活性。用含α-MEM的1%胎牛血清培养细胞24 h，然后用10 ng/ml TNF-α处理。结果TNF-α可诱导HPDL细胞MMP-1 mRNA和蛋白的表达，但其表达水平略有差异。然而，通过ELISA分析蛋白质产生没有显著差异。TNF-α诱导的ets-1 mRNA和蛋白在HPDL细胞中也无差异，而TNF-α诱导的ets-1激活则存在差异。我们正在我们的实验室进行进一步的研究，以阐明TNF-α刺激MMP-1表达变化所涉及的多态性的精确遗传分析。

Transcription factor Ets-1 play critical role in MMP-1 mRNA expression. A single nucleotide polymorphism in the promoter region of -1607 by of 1VIMP-1 have influence on Ets-1 binding activity. To clarify the ability of Ets-1 binding activity, we examined the following experiment.Methods : Human periodontal ligament cells were plated in cell culture dishes in α-MEM supplemented with 10% FBS. The confluent-stage cells were incubated with 1% FBS containing α-MEM for 24h, followed by treatment with 10 ng/ml TNF-α. After incubation, the culture supernatants and sediments were collected and analyzed by ELISA, Northern blot, and gel shift mobility assay.Results TNF-α induced MMP-1 mRNA and protein in various HPDL cells, and there were variations slightly in its expression. However, there were no significant differences in analysis of protein production by ELISA. TNF-α-induced ets-1 mRNA and protein also did not differ among HPDL cells, and Ets-lactivation in response to TNF-α showed that there were variations in activations. We are undertaking further investigations in our laboratory to elucidate the precise genetic analysis of polymorphism involved in variations of MMP-1 expression by TNF-α stimulations.