METTL14 的上调通过以 mA 依赖的方式升高 FOXO3a 的表达导致滋养层功能障碍。
Upregulation of METTL14 contributes to trophoblast dysfunction by elevating FOXO3a expression in an mA-dependent manner.
机构信息
Department of Mammary Surgery, Changzhou No.2 People's Hospital Affiliated to Nanjing Medical University, Changzhou, 213003, China.
Department of Medical Genetics, Changzhou Maternal and Child Health Care Hospital, Changzhou Medical Center, Nanjing Medical University, Changzhou, 213000, China.
出版信息
Placenta. 2022 Jun 24;124:18-27. doi: 10.1016/j.placenta.2022.05.008. Epub 2022 May 14.
INTRODUCTION
Preeclampsia, a specific complication of pregnancy, is a leading cause of perinatal and maternal mortality worldwide. N-methyladenosine (mA) is a prevalent and reversible modification of mammalian mRNAs, and is known to play an important role in various physiological and pathological processes. However, little is known about its possible effects on trophoblasts in preeclampsia.
METHODS
Colorimetric RNA mA methylation quantification assay and dot blotting were used to assess the levels of global RNA mA modification in placental tissues collected from females with normal pregnancy and preeclampsia, while the mRNA levels of major mA methyltransferases/demethylases were investigated by quantitative real-time polymerase chain reaction. The effects of methyltransferase-like 14 (METTL14) on trophoblasts were evaluated using cell counting kit-8, transwell invasion assay, autophagic flux assay, and Annexin V/propidium iodide apoptosis assay. The molecular mechanism underlying the regulation of forkhead box O3a (FOXO3a) expression by METTL14 was determined using methylated RNA immunoprecipitation and transcription inhibition assays.
RESULTS
Global RNA mA methylation and METTL14 expression were significantly increased in placental tissues obtained from patients with preeclampsia. In vitro studies showed that overexpression of METTL14 in HTR-8/SVneo cells inhibited trophoblast proliferation and invasion, but induced trophoblast autophagy and apoptosis. We further demonstrated that METTL14 epigenetically elevated FOXO3a expression via an mA-dependent mechanism. FOXO3a inhibition effectively prevented the impairment of trophoblast proliferation and invasion, and diminished the induction of trophoblast autophagy and apoptosis in METTL14-overexpressing HTR-8/SVneo cells.
DISCUSSION
Increased METTL14-mediated mA modification results in an adverse impact on trophoblast function by elevating FOXO3a expression.
简介
子痫前期是一种妊娠特有的并发症，是全球围产期和孕产妇死亡的主要原因。N6-甲基腺苷（m6A）是哺乳动物 mRNA 中一种普遍存在且可逆转的修饰方式，已知其在多种生理和病理过程中发挥着重要作用。然而，关于其在子痫前期滋养细胞中的可能作用知之甚少。
方法
采用比色法 RNA m6A 甲基化定量检测和斑点印迹法评估来自正常妊娠和子痫前期女性胎盘组织中总 RNA m6A 修饰水平，通过实时定量聚合酶链反应检测主要 m6A 甲基转移酶/去甲基酶的 mRNA 水平。使用细胞计数试剂盒-8、Transwell 侵袭实验、自噬通量实验和 Annexin V/碘化丙啶凋亡实验评估甲基转移酶样蛋白 14（METTL14）对滋养细胞的影响。采用甲基化 RNA 免疫沉淀和转录抑制实验确定 METTL14 调控叉头框蛋白 O3a（FOXO3a）表达的分子机制。
结果
子痫前期患者胎盘组织中总 RNA m6A 甲基化和 METTL14 表达显著增加。体外研究表明，在 HTR-8/SVneo 细胞中过表达 METTL14 可抑制滋养细胞增殖和侵袭，但诱导滋养细胞自噬和凋亡。我们进一步证实，METTL14 通过 m6A 依赖性机制表观遗传地上调 FOXO3a 表达。FOXO3a 抑制可有效防止 METTL14 过表达对滋养细胞增殖和侵袭的损害，并减少滋养细胞自噬和凋亡的诱导。
讨论
增加的 METTL14 介导的 m6A 修饰通过升高 FOXO3a 表达对滋养细胞功能产生不利影响。
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