METTL3通过调控miR-146a促进铁死亡水平介导双酚A导致胎盘毒性

METTL3 mediated bisphenol A(BPA)-induced placental toxicity through regulating miR-146a to promote ferroptosis

目的通过基因表达数据库(GEO)分析双酚A(BPA)致胎盘毒性的差异表达miRNA,寻找差异表达miRNA基因调控的功能通路。同时,基于基因表达数据库(GEO)分析结果,探讨METTL3调控BPA导致胎盘毒性的作用机制。方法使用GEO在线分析工具(GEO2R)对BPA暴露后差异表达基因进行筛选,对通路富集情况进行分析,使用miRNAcode和Targetscan预测软件分析BPA暴露后差异表达miRNA的靶基因。不同浓度BPA染毒JEG3细胞建立细胞模型,通过MTT检测BPA的细胞毒性,从而选择BPA浓度探讨BPA导致胎盘毒性的作用机制。通过免疫荧光检测BPA染毒后铁死亡关键蛋白GPX4分析铁死亡水平的变化;Western blotting检测BPA染毒后METTL3蛋白表达水平。结果GEO分析结果显示在BPA暴露的孕妇胎盘中高表达的miR-146a,根据GO功能分析发现多个显著的富集条目,包括细胞死亡方式、代谢以及细胞间的信息交流等。通过miRNAcode和Targetscan预测软件以miR-146a为核心预测其靶基因,预测结果发现miR-146a有20种靶基因,其中SLC7A11不仅是miR-146a的靶基因还是细胞铁死亡的关键蛋白。同时,miR-146a的表达受到m6A甲基转移酶METTL3的调控。随着BPA浓度的增加,METTL3的表达也呈现上升趋势,提示BPA通过调控METTL3的表达从而促进miR-146a的表达。结论BPA暴露通过调控METTL3促进miR-146a的表达,miR-146a通过靶向结合其靶基因SLC7A11促进细胞铁死亡,从而介导BPA导致胎盘毒性。

Objective Based on the differentially expressed miRNAs of bisphenol A(BPA)-induced toxicity through Gene Expression Database(GEO),the functional pathway of differentially expressed genes was investigated.At the same time,the mechanism of METTL3-mediated BPA-induced placental toxicity was explored.Methods GEO online analysis tool(GEO2R)was used to screen the differentially expressed genes,and the pathway enrichment was analyzed after BPA exposure.miRNAcode and Targetscan predictive software were used to identify the target genes of differentially expressed miRNA.The JEG3 cells exposed to BPA was established at different concentrations,and the cytotoxicity of BPA was detected by MTT.The concentrations of BPA were selected for the mechanism of BPA-induced placental toxicity.GPX4,the key protein of ferroptosis,was analyzed by immunofluorescence analysis.The expression level of METTL3 protein was detected by Western blotting.Results miR-146a was differentially expressed in the placenta of pregnant women exposed to BPA by GEO analysis,and significant enrichment items were obtained by GO function analysis,including cell death pattern,metabolism,and information communication between cells.miRNAcode and Targetscan analysis showed the co-network of miR-146a,which included 20 target genes.SLC7A11,a target gene of miR-146a,was not only the target gene of miR-146a but also a key protein in ferroptosis.miR-146a expression is regulated by the m6A methyltransferase METTL3.With the increase of BPA concentration,the expression of METTL3 also showed an upward trend.Conclusion BPA exposure upregulated the expression of miR-146a by regulating METTL3,and miR-146a promotes ferroptosis by targeting its target gene SLC7A11,thereby mediating the placental toxicity induced by BPA.