Gene Expression Analysis of Human Endometrial Endothelial Cells Exposed to Bisphenol A

The endocrine disrupting chemical bisphenol A (BPA) can affect reproductive organs, tissues and cells in several species. Treatment of human endometrial endothelial cells (HEECs) with 50 µM BPA decreased their proliferation compared with the control. Microarray analyses revealed that BPA affected biological processes such as the cell cycle, cell division, and cytoskeleton organization, confirming the results of the proliferation assay. Expression of three of the most differentially expressed genes identified in the microarray analysis was verified by real-time quantitative reverse transcription polymerase chain reaction in five HEEC cultures obtained from women in the proliferative phase and in five cultures obtained from women in the secretory phase of the menstrual cycle after treatment with BPA. The present study supports our previous findings of decreased proliferation and increased cell death in response to BPA, and may offer important clues to the mechanisms of action of BPA. Keywords: Exposure analysis Two-condition experiments, BPA-exposed vs. control HEEC. Biological replicates: 5 HEEC cultures, independently grown, exposed and harvested. One replicate per array. Dye-swaps for exposed and reference samples between replicates.

内分泌干扰化学物质双酚A（bisphenol A，BPA）可对多个物种的生殖器官、组织及细胞产生影响。采用50 μM浓度的BPA处理人子宫内膜内皮细胞（human endometrial endothelial cells，HEECs）后，其增殖能力较对照组出现显著下降。基因芯片（microarray）分析结果显示，BPA可调控细胞周期、细胞分裂、细胞骨架组织等生物学过程，这一发现验证了增殖实验的结果。针对基因芯片分析中筛选得到的3个差异表达最为显著的基因，本研究通过实时定量反转录聚合酶链反应（real-time quantitative reverse transcription polymerase chain reaction），对取自月经周期增殖期女性的5株HEEC培养物、以及取自分泌期女性的5株HEEC培养物分别进行BPA处理后，验证了这些基因的表达水平。本研究验证了团队此前关于BPA可引发细胞增殖能力下降、细胞死亡水平升高的研究结论，同时可为揭示BPA的作用机制提供重要线索。
关键词：暴露分析、双条件实验（BPA暴露组vs.对照组HEEC）
生物学重复：共5株HEEC培养物，均为独立培养、处理并收获的样本；每块基因芯片对应1个重复样本；各重复样本的暴露组与参照组样本间均完成了荧光染料互换。