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

转录因子Ets-1在基质金属蛋白酶-1mRNA的表达中起关键作用。1VIMP-1启动子区域-1607位单核苷酸多态对Ets-1结合活性有影响。为了阐明Ets-1结合活性的能力，我们进行了以下实验。方法：将人牙周膜细胞接种在添加10%胎牛血清的α-MEM细胞培养皿中。将融合期细胞与含有α-MEM的1%胎牛血清孵育24小时，然后用10 ng/ml肿瘤坏死因子-α处理。结果肿瘤坏死因子α可诱导HPDL细胞表达基质金属蛋白酶1m RNA和蛋白，但表达水平略有变化。而用酶联免疫吸附分析法检测蛋白质产量无显著差异。肿瘤坏死因子-α诱导的Ets-1mRNA和蛋白在不同的HPDL细胞中也没有差异，并且Ets-α对Ets-1mRNA和蛋白的激活也有差异。我们正在实验室进行进一步的研究，以阐明通过肿瘤坏死因子-α刺激引起基质金属蛋白酶-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.