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双酚A暴露后稀有鮈鲫性腺中两个类固醇生成基因的DNA甲基化状态改变

The DNA methylation status alteration of two steroidogenic genes in gonads of rare minnow after bisphenol A exposure.

作者信息

Zhang Ting, Liu Yan, Chen Hong, Gao Jiancao, Zhang Yingying, Yuan Cong, Wang Zaizhao

机构信息

College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, China.

College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, China.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2017 Aug;198:9-18. doi: 10.1016/j.cbpc.2017.05.001. Epub 2017 May 10.

Abstract

Both cytochrome P450c17 (CYP17A1) and P-450 side chain cleavage (CYP11A1) play important roles in steroid biosynthesis. According to our previous studies, bisphenol A (BPA) could regulate the mRNA expression of cyp17a1 and cyp11a1 in rare minnow Gobiocypris rarus. However, the potential mechanism of the regulation is barely understood. In the present study, aiming to explore how BPA affects the mRNA expression of cyp17a1 and cyp11a1 in testes and ovaries of G. rarus, we firstly cloned 340-bp fragment of 5' flanking region of cyp11a1 and then detected the methylation level of CpG loci involved in 5' flanking of cyp11a1 and cyp17a1 and their mRNA expression levels. Results showed that exposure to BPA significantly increased serum estradiol (E2) and 11-ketotesterone (11-KT) concentrations. Ovarian mRNA expression of cyp17a1 and cyp11a1 were significantly decreased after BPA exposure 7- for and 14-days. However, transcriptions of testicular cyp17a1 and cyp11a1 were significantly increased and decreased respectively after BPA treatment for 14days. The DNA methylation levels of cyp17a1 were decreased in ovaries on day 7 and increased in ovaries and decreased in testes respectively on day 14. The methylation levels of cyp11a1 were increased in ovaries on day 7 and both ovaries and testes on day 14. There were a significant correlation between DNA methylation at specific CpG loci and cyp17a1 and cyp11a1 genes transcription levels. In conclusion, the CpG loci methylation in 5' flanking region appears to involve in the regulation of mRNA expression of cyp17a1 and cyp11a1 mediated by BPA.

摘要

细胞色素P450c17(CYP17A1)和P - 450侧链裂解酶(CYP11A1)在类固醇生物合成中均发挥重要作用。根据我们之前的研究,双酚A(BPA)可调节稀有鮈鲫(Gobiocypris rarus)中cyp17a1和cyp11a1的mRNA表达。然而,这种调节的潜在机制几乎不为人知。在本研究中,为了探究BPA如何影响稀有鮈鲫睾丸和卵巢中cyp17a1和cyp11a1的mRNA表达,我们首先克隆了cyp11a1 5'侧翼区的340 bp片段,然后检测了cyp11a1和cyp17a1 5'侧翼区涉及的CpG位点的甲基化水平及其mRNA表达水平。结果表明,暴露于BPA显著增加了血清雌二醇(E2)和11 - 酮睾酮(11 - KT)的浓度。BPA暴露7天和14天后,卵巢中cyp17a1和cyp11a1的mRNA表达显著降低。然而,BPA处理14天后,睾丸中cyp17a1的转录显著增加,而cyp11a1的转录显著降低。cyp17a1的DNA甲基化水平在第7天卵巢中降低,在第14天卵巢中升高而在睾丸中降低。cyp11a1的甲基化水平在第7天卵巢中升高,在第14天卵巢和睾丸中均升高。特定CpG位点的DNA甲基化与cyp17a1和cyp11a1基因转录水平之间存在显著相关性。总之,5'侧翼区的CpG位点甲基化似乎参与了BPA介导的cyp17a1和cyp11a1 mRNA表达的调节。

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