人前列腺特异抗原基因启动子中一个调节序列的初步分析

PRIMARY ANALYSIS OF A REGULATORY SEQUENCE IN THE PROMOTER REGION OF HUMAN PROSTATE SPECIFIC ANTIGEN GENE

人前列腺特异抗原（ＰＳＡ）基因受雄激素调节，其雄激素应答元件（ＡＲＥ）位于－１７０附近。为确定雄激素对该基因的诱导作用是否受ＡＲＥ上游序列的影响，把ＰＳＡ启动子不同长度的ＤＮＡ片段与无启动子的ＣＡＴ报告基因相连，然后与雄激素受体表达质粒共转染人前列腺细胞ＰＣ－３。结果表明：ＡＲＥ上游（－４０６～－３７１）的一段３６ｂｐ的ＲＦ３６序列可促进雄激素对ＰＳＡ基因的诱导作用。进一步用该ＤＮＡ片段和ＰＣ－３细胞核蛋白进行区带转移测定，发现ＰＣ－３细胞中某些调节蛋白可与该序列结合，暗示该蛋白可能通过与雄激素受体的相互作用影响雄激素对ＰＳＡ启动子的诱导作用。

The human prostate specific antigen (PSA) gene has been shown to be regulated by androgen, and its androgen response element (ARE) has been determined at about 170. To determine whether the androgen induction of the gene was affected by upstream sequence of ARE, different PSA promoter DNA fragments was linked to promoterless CAT reporter gene and cotransfected with a androgen receptor expression vector in an human prostate cell line, PC 3. Transfection results indicated that a 36bp RF36 sequence (-406￣-371) at upstream of ARE could promote androgen induction of PSA gene. Further, this sequence and nucleic extract from PC 3 cells were used for band shift assay. It was found that some regulatory protein could bind to this sequence, which suggested that the protein may affect androgen induction of PSA promoter via interaction with androgen receptor.