Department of Obstetrics and Gynecology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
Canada-China-New Zealand Joint Laboratory of Maternal and Fetal Medicine, Chongqing Medical University, Chongqing, China.
Mol Med. 2022 Aug 8;28(1):92. doi: 10.1186/s10020-022-00522-4.
The forkhead box O3a protein (FoxO3a) has been reported to be involved in the migration and invasion of trophoblast, but its underlying mechanisms unknown. In this study, we aim to explore the transcriptional and metabolic regulations of FoxO3a on the migration and invasion of early placental development.
Lentiviral vectors were used to knock down the expression of FoxO3a of the HTR8/SVneo cells. Western blot, matrigel invasion assay, wound healing assay, seahorse, gas-chromatography-mass spectrometry (GC-MS) based metabolomics, fluxomics, and RNA-seq transcriptomics were performed.
We found that FoxO3a depletion restrained the migration and invasion of HTR8/SVneo cells. Metabolomics, fluxomics, and seahorse demonstrated that FoxO3a knockdown resulted in a switch from aerobic to anaerobic respiration and increased utilization of aromatic amino acids and long-chain fatty acids from extracellular nutrients. Furthermore, our RNA-seq also demonstrated that the expression of COX-2 and MMP9 decreased after FoxO3a knockdown, and these two genes were closely associated with the migration/invasion progress of trophoblast cells.
Our results suggested novel biological roles of FoxO3a in early placental development. FoxO3a exerts an essential effect on trophoblast migration and invasion owing to the regulations of COX2, MMP9, aromatic amino acids, energy metabolism, and oxidative stress.
叉头框 O3a 蛋白(FoxO3a)已被报道参与滋养细胞的迁移和侵袭,但其潜在机制尚不清楚。在本研究中,我们旨在探讨 FoxO3a 对早期胎盘发育中滋养细胞迁移和侵袭的转录和代谢调控。
使用慢病毒载体敲低 HTR8/SVneo 细胞中的 FoxO3a 表达。进行 Western blot、基质胶侵袭实验、划痕愈合实验、海瑟夫(seahorse)、基于气相色谱-质谱联用(GC-MS)的代谢组学、通量组学和 RNA 测序转录组学分析。
我们发现 FoxO3a 耗竭抑制了 HTR8/SVneo 细胞的迁移和侵袭。代谢组学、通量组学和海瑟夫实验表明,FoxO3a 敲低导致有氧呼吸向无氧呼吸转变,并增加了对细胞外营养物质中芳香族氨基酸和长链脂肪酸的利用。此外,我们的 RNA 测序还表明,FoxO3a 敲低后 COX-2 和 MMP9 的表达降低,这两个基因与滋养细胞迁移/侵袭过程密切相关。
我们的研究结果表明 FoxO3a 在早期胎盘发育中具有新的生物学作用。FoxO3a 通过调节 COX2、MMP9、芳香族氨基酸、能量代谢和氧化应激,对滋养细胞的迁移和侵袭发挥重要作用。