Zhang Caili, Li Jie, Wang Luwen, Yang Peifeng, Luo Xiaohua
Department of Obstetrics and Gynecology, The Third Affiliated Hospital of Zhengzhou University or Maternal and Child Hospital of Henan Province, No.10, Kangfuqian Street, Zhengzhou City, 450001, Henan, China.
Sci Rep. 2025 Mar 1;15(1):7345. doi: 10.1038/s41598-025-91233-3.
Preeclampsia (PE) is a pregnancy disease characterized by insufficient invasion and growth of trophoblast cells. adeno-associated virus encoding alkB homolog 1 (ALKBH1) is a demethylase in 5-methylcytosine (m5C) methylation modification. This study was performed to explore the role of ALKBH1 in hypoxia treated human extravasated trophoblast cells. Hypoxia treated human extravasated trophoblast cells (HTR-8/SVneo) was used to simulate the occurrence of PE in vitro. The cells phenotype was detected by CCK-8 and Transwell assays. The m5c levels and m5C levels of PSMD14 were analyzed by m5C dot blot and M5C Me-RIP assays. Then, the interaction between ALKBH1 and PSMD14 were confirmed by RIP and dual-luciferase reporter assays. ALKBH1 was up-regulated in hypoxia treated HTR-8/SVneo cells. Additionally, ALKBH1 knockdown increased the m5C contents, cell viability, migration and invasion abilities of hypoxia treated HTR-8/SVneo cells. Furthermore, ALKBH1 knockdown increased the m5C and mRNA levels, and mRNA stability of PSMD14. RIP and dual-luciferase reporter assays demonstrated that ALKBH1 interacted with PSMD14. Besides, PSMD14 knockdown reversed the effects of ALKBH1 silencing on cell viability, migration and invasion abilities of hypoxia treated HTR-8/SVneo cells. ALKBH1 mediated m5C levels were decreased in the hypoxia treated HTR-8/SVneo cells, which further decreased the cell viability, migration and invasion abilities through targeting the PSMD14 levels.
子痫前期（PE）是一种以滋养层细胞浸润不足和生长异常为特征的妊娠疾病。编码alkB同源物1（ALKBH1）的腺相关病毒是5-甲基胞嘧啶（m5C）甲基化修饰中的一种去甲基化酶。本研究旨在探讨ALKBH1在缺氧处理的人外渗滋养层细胞中的作用。使用缺氧处理的人外渗滋养层细胞（HTR-8/SVneo）在体外模拟PE的发生。通过CCK-8和Transwell实验检测细胞表型。通过m5C斑点杂交和M5C Me-RIP实验分析PSMD14的m5c水平和m5C水平。然后，通过RIP和双荧光素酶报告基因实验证实ALKBH1与PSMD14之间的相互作用。在缺氧处理的HTR-8/SVneo细胞中ALKBH1上调。此外，敲低ALKBH1增加了缺氧处理的HTR-8/SVneo细胞的m5C含量、细胞活力、迁移和侵袭能力。此外，敲低ALKBH1增加了PSMD14的m5C和mRNA水平以及mRNA稳定性。RIP和双荧光素酶报告基因实验表明ALKBH1与PSMD14相互作用。此外，敲低PSMD14可逆转ALKBH1沉默对缺氧处理的HTR-8/SVneo细胞的细胞活力、迁移和侵袭能力的影响。在缺氧处理的HTR-8/SVneo细胞中，ALKBH1介导的m5C水平降低，这通过靶向PSMD14水平进一步降低了细胞活力、迁移和侵袭能力。
