Abstract：
The placenta is a vital fetal organ that plays an important role in maintaining fetal sex hormone homeostasis. Xenobiotics can alter placental sex-steroidogenic enzymes and transporters, including enzymes such as aromatase (CYP19A1) and the hydroxysteroid dehydrogenases (HSDs) but studying how compounds disrupt in vivo placental metabolism is complex. Utilizing high-throughput in vitro models is critical to predict the disruption of placental sex-steroidogenic enzymes and transporters, particularly by drug candidates in the early stages of drug discovery. JAR and JEG-3 cells are the most common, simple, and cost-effective placental cell models that are capable of high-throughput screening, but how well they express the sex-steroidogenic enzymes and transporters is not well known. Here, we compared the proteomes of JAR and JEG-3 cells in the presence and absence of physiologically relevant concentrations of dehydroepiandrosterone (DHEA, 8 µM) and testosterone (15 nM) to aid the characterization of sex-steroidogenic enzymes and transporters in these cell models. Global proteomics analysis detected 2931 and 3449 proteins in JAR cells and JEG-3 cells, respectively. However, dramatic differences in sex-steroidogenic enzymes and transporters were observed between these cells. In particular, the basal expression of steroid sulfatase (STS), HSD17B1, and HSD17B7 were unique to JEG-3 cells. JEG-3 cells also showed significantly higher protein levels of aldo-keto reductase (AKR) 1A1 and AKR1B1, while JAR cells showed significantly higher levels of HSD17B4 and HSDB12. Aldehyde dehydrogenase (ALDH) 3A2 and HSD17B11 enzymes as well as the transporters sterol O-acyltransferase (SOAT) 1 and ATP binding cassette subfamily G2 (ABCG2) were comparable between the cell lines, whereas sulfotransferases (SULTs) were uniquely present within JAR cells. Androgen treatments significantly lowered HSD17B11, HSD17B4, HSD17B12, and ALDH3A2 levels in JAR cells. DHEA treatment significantly raised the level of HSD17B1 by 51 % in JEG-3 cells, whereas CYP19A1 was increased to significant levels in both JAR and JEG-3 cells after androgen treatments. The proteomics data were supported by a complementary targeted metabolomics analysis of culture media in the DHEA (8 µM) and testosterone (15 nM) treated groups. This study has indicated that untreated JEG-3 cells express more sex-steroidogenic enzymes and transporters. Nevertheless, JEG-3 and JAR cells are unique and their respective proteomics data can be used to select the best model depending on the hypothesis.
摘要：
胎盘是重要的胎儿器官，在维持胎儿性激素稳态中起着重要作用。异种生物可以改变胎盘性类固醇生成酶和转运蛋白，包括芳香酶（CYP19A1）和羟基类固醇脱氢酶（HSD）等酶，但研究化合物如何破坏体内胎盘代谢是复杂的。利用高通量体外模型对于预测胎盘性促类固醇酶和转运蛋白的破坏至关重要。特别是在药物发现的早期阶段的候选药物。JAR和JEG-3细胞是最常见的，简单,和具有成本效益的胎盘细胞模型，能够进行高通量筛选，但是它们表达性类固醇酶和转运蛋白的程度尚不清楚。这里,我们比较了在存在和不存在生理相关浓度的脱氢表雄酮（DHEA，8µM）和睾丸激素（15nM），以帮助表征这些细胞模型中的性类固醇生成酶和转运蛋白。全球蛋白质组学分析在JAR细胞和JEG-3细胞中检测到2931和3449蛋白，分别。然而,在这些细胞之间观察到性促类固醇酶和转运蛋白的巨大差异。特别是,类固醇硫酸酯酶（STS）的基础表达，HSD17B1和HSD17B7对于JEG-3细胞是独特的。JEG-3细胞还显示出明显较高的醛酮还原酶（AKR）1A1和AKR1B1蛋白水平，而JAR细胞显示出明显较高的HSD17B4和HSDB12水平。乙醛脱氢酶（ALDH）3A2和HSD17B11酶以及转运蛋白固醇O-酰基转移酶（SOAT）1和ATP结合盒亚家族G2（ABCG2）在细胞系之间具有可比性，而磺基转移酶（SULTs）在JAR细胞中独特存在。雄激素处理显著降低JAR细胞中HSD17B11、HSD17B4、HSD17B12和ALDH3A2水平。DHEA处理显著提高了JEG-3细胞中HSD17B1的水平51％，而在雄激素处理后，CYP19A1在JAR和JEG-3细胞中均显著升高。蛋白质组学数据得到DHEA(8μM)和睾酮(15nM)处理组中培养基的互补靶向代谢组学分析的支持。这项研究表明，未经处理的JEG-3细胞表达更多的性类固醇生成酶和转运蛋白。然而，JEG-3和JAR细胞是独特的，它们各自的蛋白质组学数据可用于根据假设选择最佳模型。
