中文摘要：

目的研究17β-雌二醇对解聚素金属蛋白酶9（ADAM9）表达水平的影响。方法应用盐析法提取人类基因组DNA，扩增ADAM9基因启动子目的片断，构建表达ADAM9启动子的报告基因质粒。应用脂质体转染法，分别转染人人神经母细胞瘤细胞SK．N—SH、人胚肾细胞HEK293和宫颈癌细胞HeLa内。17β-雌二醇处理各组细胞后，检测ADAM9启动子转录效率。结果成功扩增了ADAM9启动子区1676bp目的片段，构建了ADAM9启动子报告基因质粒pGL．ADAM9，瞬时转染入细胞内。17β-雌二醇处理SK_N．SH细胞后，ADAM9启动子转录活性上升至31．250＋1．328，较处理前的15．471＋1．256增加约2倍，差异有统计学意义（P〈0．05）。HEK293细胞内，ADAM9启动子转录活性上升至13．499＋0．866，较处理前的8．513＋1．356增加约1．6倍，差异有统计学意义（氏0．05）。HeLa细胞中没有观察到1713．雌二醇对ADAM9启动子转录活性的影响。结论17B．雌二醇能上调人ADAM9启动子的表达水平，是ADAM9基因表达的激活剂。

英文摘要：

Objective To study the effect of 17[3-estradiol on the a-disintegrin and metalloprotease 9 （ADAM9） expression. Methods Genomic DNA was prepared from human blood sample using salting-out method. Systematic screening of ADAM9 promoter region was performed using standard PCR and direct sequencing. The reporter plasmid expressed ADAM9 promoter was constructed and transiently transfected into SK-N-SH （human neuroblastoma cells）, HEK293 （human embryonic kidney cells） and HeLa （cervical cancer cells） using liposome transfection method. Transfected cells were treated with 1713-estradiol for 24 h, and then, they were lysed and assayed to detect the transcriptional efficiency of ADAM9 promoter by dual-luciferase reporter assay system. Results We cloned the 1676 fragment of the proximal promoter of ADAM9 into the upstream of luciferase reporter gene （pGL-ADAM9）. They were transiently transfected into the SK-N-SH, HEK293 and HeLa cells. By 17[3-estradiol treatment, the ADAM9 transcriptional activities were significantly increased： the ADAM9 transcriptional activities in SK-N-SH cells increased about two times from （15.471_＋1.256） before treatment to （31.250_＋1.328） after treatment, and the ADAM9 transcriptional activities in HEK293 cells increased about 1.6 times from （8.513_＋1.356） before treatment to （13.499_＋0.866） after treatment （P〈 0.05）; however, no effect of 1713-estradiol on ADAM9 transcriptional activities in HeLa cells was noted. Conclusion Estrogen can increase the expression of ADAM9 promoter and can act as an activator of human A DA M9 promoter activity.