Chen Chao, Gao Jing, Chen Dan, Liu Jinyu, He Biwei, Chen Yan, Zhang Huijuan, Yang Xingyu, Cheng Weiwei
International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Shanghai Key Laboratory of Embryo Original Disease, Shanghai, China.
Cell Cycle. 2022 Dec;21(23):2517-2532. doi: 10.1080/15384101.2022.2103897. Epub 2022 Jul 28.
Preeclampsia (PE) is a pregnancy-associated disease that may cause maternal and fetal morbidity and mortality. The dysregulation of microRNAs (miRNAs) and their potential functions has been an important direction for elucidating the mechanism of preeclampsia in recent years. The present study investigated whether miR-4443 was significantly increased in the placentas of severe preeclamptic patients, and the upregulation of miR-4443 inhibited the migration and invasion of HTR-8/SVneo cells according to transwell assays. Matrix metallopeptidase 2 (MMP2), which is involved in the degradation of extracellular matrix (ECM) components and harbors a miR-4443-binding site within its 3'-UTR as confirmed by a luciferase reporter assay, was identified to be directly inhibited by miR-4443. Moreover, siRNA targeting MMP2 imitated the effects of overexpressed miR-4443 on HTR-8/SVneo cell invasion and migration, whereas rescue experiments showed that MMP2 reversed this inhibitory function of miR-4443. Heparin-binding EGF-like growth factor (HB-EGF), as the downstream gene of MMP2, plays an important role in trophoblast invasion, and we confirmed that the expression of HB-EGF/EGFR pathway-related biomolecules was consistent with MMP2 influenced by upregulating and downregulating miR-4443 and that activated EGFR further transmitted intracellular downstream signaling via the MAPK pathway according to western blot assay. In conclusion, we demonstrated that miR-4443 suppresses the migration and invasion of trophoblasts, and its inhibitory effects are at least partially mediated by the suppression of MMP2. This inhibition might further affect the progression of preeclampsia through the HB-EGF/EGFR pathway, thus providing a new clue on the role of miR-4443 in the pathogenesis of preeclampsia.
子痫前期(PE)是一种与妊娠相关的疾病,可能导致母婴发病和死亡。近年来,微小RNA(miRNA)的失调及其潜在功能一直是阐明子痫前期发病机制的重要方向。本研究调查了重度子痫前期患者胎盘组织中miR-4443是否显著上调,并且通过Transwell实验发现miR-4443的上调抑制了HTR-8/SVneo细胞的迁移和侵袭。荧光素酶报告基因实验证实,基质金属蛋白酶2(MMP2)参与细胞外基质(ECM)成分的降解,且其3'-UTR区域存在miR-4443结合位点,研究发现miR-4443可直接抑制MMP2。此外,靶向MMP2的小干扰RNA(siRNA)模拟了过表达miR-4443对HTR-8/SVneo细胞侵袭和迁移的影响,而挽救实验表明MMP2可逆转miR-4443的这种抑制作用。肝素结合表皮生长因子(HB-EGF)作为MMP2的下游基因,在滋养细胞侵袭中起重要作用,蛋白质免疫印迹实验证实,上调和下调miR-4443影响MMP2表达时,HB-EGF/表皮生长因子受体(EGFR)通路相关生物分子的表达与之一致,并且激活的EGFR通过丝裂原活化蛋白激酶(MAPK)通路进一步传递细胞内下游信号。总之,本研究表明miR-4443抑制滋养细胞的迁移和侵袭,其抑制作用至少部分是通过抑制MMP2介导的。这种抑制作用可能通过HB-EGF/EGFR通路进一步影响子痫前期的进展,从而为miR-4443在子痫前期发病机制中的作用提供了新线索。