Direct effects of diethylstilbestrol on the gene expression of the cholesterol side-chain cleavage enzyme (P450scc) in testicular Leydig cells.
Direct effects of diethylstilbestrol on the gene expression of the cholesterol side-chain cleavage enzyme (P450scc) in testicular Leydig cells.
复制标题
DOI：
10.1016/j.lfs.2010.06.020
复制
发表时间：
2010-08
期刊：
影响因子：
6.1
通讯作者：
K. Warita;T. Mitsuhashi;T. Sugawara;Y. Tabuchi;Takashi Tanida;Zhi-yu Wang;Y. Matsumoto;T. Yokoyama-T.-Yok
中科院分区：
文献类型：
--
作者：
K. Warita;T. Mitsuhashi;T. Sugawara;Y. Tabuchi;Takashi Tanida;Zhi-yu Wang;Y. Matsumoto;T. Yokoyama-T.-Yok
目的研究雌激素性内分泌干扰物作用的确切机制，我们评估了合成雌激素己烯雌酚（DES）对Leydig细胞类固醇合成的直接影响，特别强调了胆固醇侧链裂解酶P450 SCC的表达。此外，DES的作用机制与内源性雌激素17β-雌二醇（E2）的作用机制进行了比较，后者具有与DES相当的效力。主要方法1用5×10−8μM至5μM DES或E2处理Leydig细胞24 h，和P450 SCC基因表达和组蛋白修饰的转录激活用逆转录-聚合酶链反应检测（DES处理的和E2处理的细胞中SP 450 SCC mRNA的表达分别与DES和E2的剂量成反比地降低;然而，cAMP刺激诱导表达恢复到与对照组大致相等的水平。在DES处理的细胞中，ChIP检测显示P450 SCC启动子区域的组蛋白去乙酰化。有趣的是，E2没有引起组蛋白去乙酰化。意义在类固醇生成的早期阶段，DES和E2直接诱导P450 SCC mRNA表达的减少，与其剂量成反比，和治疗与cAMP恢复减少的P450 SCC mRNA表达。此外，DES可诱导P450 SCC基因组蛋白修饰的改变，天然雌激素和合成雌激素化合物如DES可通过不同的分子机制诱导生殖障碍。
AIMSTo investigate the precise mechanisms underlying the action of estrogenic endocrine disruptors, we evaluated the direct effects of synthetic estrogen diethylstilbestrol (DES) on steroidogenesis in Leydig cells, with particular emphasis on the expression of the cholesterol side-chain cleavage enzyme P450scc. Furthermore, the mechanism underlying the action of DES was compared with that of endogenous estrogen 17β-estradiol (E2), which has a potency equivalent to that of DES.MAIN METHODSTTE1 Leydig cells were treated with 5×10−8μM to 5μM DES or E2 for 24h, and P450scc gene expression and the histone modifications underlying their transcriptional activation were examined using reverse transcription-polymerase chain reaction (RT-PCR) and chromatin immunoprecipitation (ChIP), respectively.KEY FINDINGSP450scc mRNA expression in the DES-treated and E2-treated cells reduced in inverse proportion to the dose of DES and E2, respectively; however, cAMP stimulation induced a recovery in the expression to a level approximately equal to those in the controls. In the DES-treated cells, ChIP assay revealed histone deacetylation in the P450scc promoter region. Interestingly, E2 did not cause histone deacetylation.SIGNIFICANCEIn the early stages of steroidogenesis, DES and E2 directly induced a reduction in P450scc mRNA expression in inverse proportion to their doses, and treatment with cAMP restored the decreased P450scc mRNA expression. Furthermore, DES can induce alterations in the histone modification of the P450scc gene, and natural estrogen and synthetic estrogenic compounds such as DES may induce reproductive disorders through different molecular mechanisms.