雄激素应答元件假冒DNA对PSA基因启动子的抑制作用

研究雄激素应答元件假冒DNA(AREdecoy)对前列腺特异抗原 (PSA)基因启动子的抑制作用 .联合运用报告基因和假冒DNA策略 ,构建了含PSA基因 5′侧启动子区 6 40bpDNA的萤光素酶表达载体pGL3 PSA ,与人工合成的双链硫代ARE假冒DNA共转染前列腺癌细胞株PC3 M并作用不同的时间 (2 4h、4 8h、72h) .应用双萤光素酶测定系统 ,检测萤光素酶的表达活性 .结果显示 :AREdecoyDNA显著抑制报告基因萤光素酶的表达 ,抑制率可达 95 % ,而对照decoyDNA无此作用 .作用不同的时间对萤光素酶活性的抑制无显著性差异

Inhibition of Androgen Responsive Element Decoy DNA on the Promoter of Prostate Specific Antigen

To observe the effect of exogenous androgen responsive element decoy on the promoter of prostate specific antigen (PSA), pGL3 PSA luciferase expression vector containing 640 bp DNA fragment of the PSA gene 5′ promoter region was constructed. Meanwhile, a 23 mer phosphorothioated ARE decoy based on the deduced ARE sequence at the promoter region of PSA gene was synthesized. pGL3 PSA plasmid and the ARE decoy DNA were cotransfected into PC3 M cell by lipofectamine TM 2000. Through detecting the activity of luciferase, the effect of ARE decoy on the promoter of PSA was studied. The results showed that the activity of luciferase was significantly reduced in the ARE decoy transfected cells, though not in the cells transfected with the control decoy. There were no significant differences in the effect of ARE decoy in different time.