5α-双氢睾酮对解聚素金属蛋白酶9表达水平的影响

The effect of 5α-dihydrotestosterone on the expression of a disintegrin and metalloprotease 9

目的研究5α-双氢睾酮(5α-DHT)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基因启动子区目的片断,构建表达ADAM9启动子的报告基因质粒。应用脂质体转染法,分别转染入人神经母细胞瘤细胞(SH-SY5Y)、人前列腺癌细胞(LNCaP)和人宫颈癌上皮细胞(HeLa)内。5α-DHT处理各组细胞后,检测ADAM9启动子转录效率。结果我们扩增了ADAM9启动子区1676bp目的片断,构建了ADAM9启动子报告基因质粒pGL-ADAM9,将其瞬时转染入上述细胞内。5α-DHT干预细胞后,ADAM9启动子转录活性与非处理组相比差异无统计学意义(P>0.05)。结论上述三种细胞中,5α-DHT对ADAM9启动子表达水平无显著影响。

Objective To study the effect of 5α-dihydrotestosterone （5α-DHT） on the expression of a disintegrin and metalloprotease 9 （ADAM9） which is one of o^-secretases. Methods Genomic DNA was prepared from human blood sample using the salting-out method. Systematic screening of ADAM9 promoter region was performed using standard PCR and direct sequencing. The reporter plasmid which expressed the promoter region of ADAM9 was constructed and transiently transfected into SH-SYSY, LNCaP and HeLa cells using lipofeetamine 2000. Transfected cells were treated under 5α-DHT, then they were lysed and assayed by Dual-Luciferase reporter assay system.Results The 1676 fragment of the proximal promoter of ADAM9 were cloned into upstream of the luciferase reporter gene （pGL-ADAM9）. They were transiently transfected into cells. Under 5α-DHT treatment, the ADAM9 transcriptional activities were not significantly increased than the control groups （P 〉 0.05 ）. Conclusion In SH-SYSY, LNCaP and HeLa cells, 5α-DHT can not increase the expression of oL-secretase ADAM9.