Department of Endocrinology, Institute of Zoology, Jagiellonian University, Krakow, Poland.
Andrology. 2014 Mar;2(2):186-97. doi: 10.1111/j.2047-2927.2013.00172.x. Epub 2013 Dec 18.
In both epididymis and prostate the dynamic cross-talk between the cells is hormonally regulated and, in part, through direct cell-to-cell interactions. Functionality of the male reproductive organs may be affected by exposure to specific chemicals, so-called 'reprotoxicants'. In this study we tested whether early postnatal and prepubertal exposure to anti-androgen flutamide altered the expression of adherens junction genes encoding E-cadherin (CDH1) and β-catenin (CTNNB1) in adult pig epididymis and prostate. In addition, the expression of mRNAs and proteins for 5α-reductase (ST5AR2) and aromatase (CYP19A1) were examined to show whether flutamide alters metabolism of testosterone. Thus, flutamide was injected into male piglets between Days 2 and 10 and between Days 90 and 98 postnatally (PD2 and PD90; 50 mg/kg bw), tissues that were obtained on postnatal Day 270. To assess the expression of the genes and proteins, real-time RT-PCR and Western blot were performed respectively. Moreover, adherens junction proteins were localized by immunohistochemistry. In response to flutamide, CDH1 and CTNNB1 expressions were down-regulated along the epididymis, mostly in PD2 group (p < 0.001, p < 0.01). In the prostate, CDH1 mRNA and protein expressions were significantly down-regulated (p < 0.01), whereas CTNNB1 mRNA was slightly up-regulated in both flutamide-treated groups. CTNNB1 protein level was markedly elevated in both PD2 (p < 0.001) and PD90 (p < 0.01) groups. In the epididymis, the expression of ST5AR2 and CYP19A1 was down- and up-regulated, respectively (p < 0.05), whereas in the prostate evident decrease in CYP19A1 expression (p < 0.001, p < 0.01, p < 0.05) was demonstrated. In both tissues, membranous immunolocalization of CTNNB1 suggests its involvement in cell-cell adhesion. Overall, flutamide administration resulted in suppression of androgen action in the epididymis and prostate leading to deregulation of CDH1 and CTNNB1 gene expressions which is probably caused by the alterations in the expression of ST5AR2 and CYP19A1 in both reproductive organs.
在附睾和前列腺中，细胞之间的动态串扰受激素调节，部分受细胞间直接相互作用的调节。男性生殖器官的功能可能会受到特定化学物质（所谓的“生殖毒素”）的影响。在这项研究中，我们测试了新生后和青春期前暴露于抗雄激素氟他胺是否会改变成年猪附睾和前列腺中编码 E-钙黏蛋白（CDH1）和β-连环蛋白（CTNNB1）的黏附连接基因的表达。此外，还检查了 5α-还原酶（ST5AR2）和芳香酶（CYP19A1）的 mRNA 和蛋白质的表达，以显示氟他胺是否改变了睾酮的代谢。因此，在新生后第 2 天和第 10 天（PD2 和 PD90）以及新生后第 90 天和第 98 天（PD2 和 PD90）之间向雄性仔猪注射氟他胺（50mg/kg bw），并在新生后第 270 天获得组织。为了评估基因和蛋白质的表达，分别进行了实时 RT-PCR 和 Western blot。此外，通过免疫组织化学定位黏附连接蛋白。氟他胺处理后，CDH1 和 CTNNB1 的表达沿附睾下调，主要在 PD2 组（p < 0.001，p < 0.01）。在前列腺中，CDH1 mRNA 和蛋白质表达显著下调（p < 0.01），而 CTNNB1 mRNA 在两个氟他胺处理组中略有上调。在 PD2（p < 0.001）和 PD90（p < 0.01）组中，CTNNB1 蛋白水平明显升高。在附睾中，ST5AR2 和 CYP19A1 的表达分别下调和上调（p < 0.05），而在前列腺中，CYP19A1 的表达明显下降（p < 0.001，p < 0.01，p < 0.05）。在这两种组织中，CTNNB1 的膜免疫定位表明其参与细胞-细胞黏附。总的来说，氟他胺的给药导致附睾和前列腺中雄激素作用受到抑制，导致 CDH1 和 CTNNB1 基因表达失调，这可能是由于这两种生殖器官中 ST5AR2 和 CYP19A1 表达的改变所致。
