Mansur Abdallah, Israel Ariel, Combelles Catherine M H, Adir Michal, Racowsky Catherine, Hauser Russ, Baccarelli Andrea A, Machtinger Ronit
Infertility and IVF Unit, Department of Obstetrics and Gynecology, Chaim Sheba Medical Center and Sackler School of Medicine, Tel-Aviv University, Tel Hashomer 52561, Israel.
Department of Family Medicine, Clalit Health Services, Jerusalem, Israel.
Hum Reprod. 2017 Feb;32(2):409-417. doi: 10.1093/humrep/dew316. Epub 2016 Dec 15.
Does bisphenol-A (BPA) affect gene expression in human membrana granulosa cells (MGC)?
In vitro, short exposure to supra-physiological concentrations of BPA alters human MGC gene expression.
Exposure to BPA may interfere with reproductive endocrine signaling. In vitro studies, mostly in animal models, have shown an inverse correlation between exposure to BPA and follicular growth, meiosis, and steroid hormone production in granulosa cells.
STUDY DESIGN, SIZE, DURATION: Primary cultures of MGC obtained from 24 patients undergoing IVF (for PGD, male factor infertility or unexplained infertility) were exposed to various concentrations of BPA (0, 0.02, 0.2, 2 or 20 µg/ml) for 48 h.
PARTICIPANTS/MATERIALS, SETTING, METHODS: The study was conducted in a university-affiliated hospital. Microarray analysis was used to identify genes exhibiting expression changes following BPA exposure. Genes significantly altered were identified based on changes greater than 2-fold relative to the control group (not treated by BPA) and a Student's t-test P-value <0.05. Statistical significance was adjusted for multiple comparisons using the Benjamini-Hochberg method. Alterations in the expression of genes that are involved in the enriched functional annotations altered by BPA at the concentration of 20 µg/ml were confirmed by real-time PCR.
A distinct pattern of gene expression was observed in primary cultures of MGC exposed to the highest BPA concentration compared with untreated cells. We identified 652 genes that exhibited at least 2-fold differences in expression after BPA exposure (all P < 0.05 versus untreated). These genes were significantly enriched for annotations related to cell cycle progression, segregation of chromosomes, steroid metabolism, apoptosis, lipid synthesis, oocyte maturation and chromosomal alignment. No significant changes in gene expression were found at the lower doses of BPA most relevant to human exposure.
N/A.
LIMITATIONS, REASONS FOR CAUTION: Human exposure to BPA in vivo occurs over long periods of time. In this in vitro model, cells were exposed to the chemical for 48 h only. Thus, the effects of BPA on the human follicle might be underestimated.
As BPA exposure is ubiquitous, understanding the effects of the chemical on the ovary, specifically in women of reproductive age, has public health significance. The clinical evidence to date points to an association between BPA exposure and impaired IVF outcome, although not all studies have shown negative effects. Our study adds valuable mechanistic information showing that exposure to BPA alters granulosa cell gene expression at high and supra-physiological doses.
STUDY FUNDING/COMPETING INTERESTS: This study was supported by grant number 1936/12 from the ISF. The authors have nothing to disclose.
双酚A（BPA）是否会影响人颗粒膜细胞（MGC）中的基因表达？
在体外，短期暴露于超生理浓度的双酚A会改变人颗粒膜细胞的基因表达。
接触双酚A可能会干扰生殖内分泌信号传导。体外研究（大多在动物模型中）表明，接触双酚A与颗粒细胞中的卵泡生长、减数分裂及类固醇激素产生呈负相关。
研究设计、规模、持续时间：从24例接受体外受精（用于植入前遗传学诊断、男性因素不育或不明原因不育）的患者中获取颗粒膜细胞原代培养物，将其暴露于不同浓度的双酚A（0、0.02、0.2、2或20μg/ml）中48小时。
参与者/材料、环境、方法：该研究在一家大学附属医院进行。采用微阵列分析来鉴定双酚A暴露后表现出表达变化的基因。基于相对于对照组（未用双酚A处理）变化大于2倍以及学生t检验P值<0.05来鉴定显著改变的基因。使用Benjamini-Hochberg方法对多重比较的统计显著性进行校正。通过实时PCR确认在20μg/ml浓度下双酚A改变的参与富集功能注释的基因表达变化。
与未处理细胞相比，在暴露于最高双酚A浓度的颗粒膜细胞原代培养物中观察到明显的基因表达模式。我们鉴定出652个基因在双酚A暴露后表现出至少2倍的表达差异（与未处理组相比，所有P<0.05）。这些基因在与细胞周期进程、染色体分离、类固醇代谢、细胞凋亡、脂质合成、卵母细胞成熟和染色体排列相关的注释中显著富集。在与人类暴露最相关的较低剂量双酚A下未发现基因表达的显著变化。
无。
局限性、谨慎原因：人体在体内长期接触双酚A。在这个体外模型中，细胞仅暴露于该化学物质48小时。因此，双酚A对人卵泡的影响可能被低估。
由于双酚A暴露无处不在，了解该化学物质对卵巢的影响，特别是对育龄女性的影响，具有公共卫生意义。尽管并非所有研究都显示出负面影响，但迄今为止的临床证据表明双酚A暴露与体外受精结果受损之间存在关联。我们的研究增加了有价值的机制信息，表明在高剂量和超生理剂量下接触双酚A会改变颗粒细胞基因表达。
研究资金/利益冲突：本研究由以色列科学基金会的1936/12号资助。作者无利益冲突声明。
