Liu Yanhua, Liu Yingru, Zhang Xiaoying, Zhu Xiaoxiong, Liu Yaping
Department of Obstetrics, the Fourth Hospital of Hebei Medical University, Shijiazhuang City, Hebei Province, China.
Medical insurance office, the Fourth Hospital of Hebei Medical University, Shijiazhuang City, Hebei Province, China.
J Biochem Mol Toxicol. 2025 Sep;39(9):e70458. doi: 10.1002/jbt.70458.
Pre-eclampsia (PE) represents a serious pregnancy complication characterized by impaired trophoblast function. Although methyltransferase-like 14 (METTL14) has been implicated in PE pathogenesis and trophoblast dysfunction, its precise molecular mechanisms remain unclear. mRNA expression was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Immunoblot analysis was used for protein expression evaluation. Total N6-methyladenosine (m6A) RNA methylation was analyzed using an assay kit. Cell tube formation, invasion and colony formation were assessed by tube formation, transwell and colony formation assays, respectively. Interleukin-8 (IL-8), IL-1beta (IL-1β), and IL-6 levels were quantified by enzyme-linked immunosorbent assay (ELISA). The influence of METTL14 in forkhead box protein 1 (FOXP1) was tested by methylated RNA immunoprecipitation (MeRIP) and mRNA stabilization assays. The FOXP1/transmembrane BAX inhibitor motif-containing 6 (TMBIM6) relationship was validated by chromatin immunoprecipitation (ChIP) and luciferase assays. Total m6A methylation levels and METTL14 expression were significantly increased in placental samples of PE patients. Overexpression of METTL14 diminished cell colony formation, invasion, and tube formation abilities of human HTR-8/SVneo trophoblast cells and promoted their pro-inflammatory cytokine production. METTL14 epigenetically reduced FOXP1 expression via an m6A dependent mechanism. Re-expression of FOXP1 exerted a counteracting impact on METTL14-driven dysfunction and inflammation of HTR-8/SVneo cells. FOXP1 promoted TMBIM6 transcription, and METTL14 reduced TMBIM6 expression by FOXP1. FOXP1 regulated HTR-8/SVneo cell phenotypes and inflammation by TMBIM6. Our findings identify a new epigenetic mechanism, the METTL14/FOXP1/TMBIM6 axis, with the ability to affect trophoblast function and inflammation.
子痫前期（PE）是一种严重的妊娠并发症，其特征为滋养细胞功能受损。尽管甲基转移酶样14（METTL14）与PE发病机制及滋养细胞功能障碍有关，但其确切分子机制仍不清楚。通过定量实时聚合酶链反应（qRT-PCR）检测mRNA表达。采用免疫印迹分析评估蛋白质表达。使用检测试剂盒分析总N6-甲基腺苷（m6A）RNA甲基化。分别通过管形成、Transwell和集落形成试验评估细胞管形成、侵袭和集落形成。通过酶联免疫吸附测定（ELISA）定量白细胞介素-8（IL-8）、IL-1β和IL-6水平。通过甲基化RNA免疫沉淀（MeRIP）和mRNA稳定性试验检测METTL14对叉头框蛋白1（FOXP1）的影响。通过染色质免疫沉淀（ChIP）和荧光素酶试验验证FOXP1/含跨膜BAX抑制基序6（TMBIM6）的关系。PE患者胎盘样本中总m6A甲基化水平和METTL14表达显著增加。METTL14的过表达降低了人HTR-8/SVneo滋养细胞的细胞集落形成、侵袭和管形成能力，并促进其促炎细胞因子的产生。METTL14通过m6A依赖机制在表观遗传上降低FOXP1表达。FOXP1的重新表达对METTL14驱动的HTR-8/SVneo细胞功能障碍和炎症产生拮抗作用。FOXP1促进TMBIM6转录，而METTL14通过FOXP1降低TMBIM6表达。FOXP1通过TMBIM6调节HTR-8/SVneo细胞表型和炎症。我们的研究结果确定了一种新的表观遗传机制，即METTL14/FOXP1/TMBIM6轴，其具有影响滋养细胞功能和炎症的能力。
