Department of Obstetrics and Gynaecology, Zhongda Hospital, School of Medicine, Southeast University, Nanjing 210009, China.
Xinghua City People's Hospital, Xinghua 225700, China.
Ecotoxicol Environ Saf. 2021 Mar 15;211:111945. doi: 10.1016/j.ecoenv.2021.111945. Epub 2021 Jan 28.
The study aimed to recognize potential molecular targets and signal pathways whereby phenolic environmental estrogen promotes the proliferation of uterine leiomyoma cells.
Primary cultured cell lines of uterine leiomyoma were treated with 0.1% DMSO, 10.0μmol/L Bisphenol A (BPA), and 32.0μmol/L Nonylphenol (NP) for 48 h before RNA-seq was performed. Those genes affected by BPA and NP were identified. Then, Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and Protein-protein Interaction (PPI) analysis were performed. Quantitative real-time polymerase chain reaction (q-PCR) and western blot were used to verify the differentially expressed gene and protein.
Compared to with the control group, 739 differentially expressed genes were identified in both the BPA group and the NP group. GO enrichment analysis showed that the most enriched GO terms were connective tissue development and G1/S transition of mitotic cell cycle, and extracellular matrix. The results of KEGG enrichment analysis showed that differentially expressed mRNA were enriched mainly in three primary pathways, including environmental information processing, human diseases, and cellular processes. The cell cycle, PI3K-Akt signaling pathway are significantly enriched. The q-PCR and western blot verified the cell cycle associated genes and proteins were upregulated in both BPA group and NP group. Both BPA and NP activated the PI3K-AKT signaling pathway.
Phenolic environmental estrogens may promote the proliferation and cell cycle progression of uterine leiomyoma cells through rapid non-genomic ER signaling, which leads to disordered cell cycle regulation and accelerates the transition of the cell cycle from G0/G1 phase to S phase. In addition, as an external stimulant, phenolic estrogen promotes the upregulation of inflammatory factors in uterine leiomyomas.
本研究旨在识别酚类环境雌激素促进子宫肌瘤细胞增殖的潜在分子靶标和信号通路。
用 0.1%DMSO、10.0μmol/L 双酚 A（BPA）和 32.0μmol/L 壬基酚（NP）处理原代培养的子宫肌瘤细胞系 48 h 后，进行 RNA-seq。鉴定受 BPA 和 NP 影响的基因。然后进行基因本体论（GO）富集、京都基因与基因组百科全书（KEGG）通路富集和蛋白质-蛋白质相互作用（PPI）分析。采用定量实时聚合酶链反应（q-PCR）和蛋白质印迹法验证差异表达基因和蛋白。
与对照组相比，BPA 组和 NP 组均有 739 个差异表达基因。GO 富集分析显示，最富集的 GO 术语是结缔组织发育和有丝分裂细胞周期的 G1/S 期过渡，以及细胞外基质。KEGG 富集分析结果表明，差异表达的 mRNA 主要富集在三个主要途径中，包括环境信息处理、人类疾病和细胞过程。细胞周期、PI3K-Akt 信号通路明显富集。q-PCR 和蛋白质印迹法验证了 BPA 组和 NP 组中与细胞周期相关的基因和蛋白上调。BPA 和 NP 均激活了 PI3K-AKT 信号通路。
酚类环境雌激素可能通过快速非基因组 ER 信号促进子宫肌瘤细胞的增殖和细胞周期进程，导致细胞周期调控紊乱，加速细胞周期从 G0/G1 期向 S 期的过渡。此外，作为一种外部刺激物，酚类雌激素促进子宫肌瘤中炎症因子的上调。
