Department of Obstetrics and Gynecology, Osaka Medical and Pharmaceutical University, Osaka, Japan.
Department of Obstetrics and Gynecology, Osaka Medical and Pharmaceutical University, Osaka, Japan.
Placenta. 2022 Jun 1;123:5-11. doi: 10.1016/j.placenta.2022.04.004. Epub 2022 Apr 22.
Appropriate implantation of trophoblast cells is necessary for successful pregnancy outcome. This process requires proper migration and invasion of trophoblast cells into the maternal endometrium and the myometrium. Dysregulation of circulating microRNAs in preeclampsia has been reported in several studies. Furthermore, miR-486-5p was reportedly increased within exosomes derived from maternal circulation in preeclamptic pregnancy. However, the roles of elevated miR-486-5p in preeclampsia has not yet been clarified.
HTR8/SVneo trophoblast cells were transfected with miR-486-5p, and the ARHGAP5 expression was examined by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blotting. A reporter assay using a luciferase construct containing the ARHGAP5 3'-untranslated region (3'UTR) was performed to determine whether or not ARHGAP5 is a direct target of miR-486-5p. Changes in migration and invasion abilities were examined by a wound healing assay and invasion assay, respectively.
The ARHGAP5 expression was significantly decreased in miR-486-5p-transfected cells according to RT-qPCR and Western blotting. A dual luciferase reporter gene assay showed that miR-486-5p acts directly on the 3'UTR of ARHGAP5 mRNA. The migration and invasion abilities were suppressed in miR-486-5p-transfected cells. Downregulation of ARHGAP5 by small interfering RNA transfection inhibited trophoblast cell migration and invasion, resembling that of miR-486-5p transfection.
The migration and invasion abilities of HTR8/SVneo cells were suppressed by miR-486-5p at least partly through inhibiting the ARHGAP5 expression. These data suggest that miR-486-5p is involved in the pathogenesis of preeclampsia and that miR-486-5p is a viable potential biomarker for predicting the onset risk of preeclampsia.
适当的滋养细胞植入对于成功的妊娠结局是必要的。这个过程需要滋养细胞正确地迁移和侵入母体子宫内膜和子宫肌层。几项研究报道子痫前期患者的循环 microRNAs 存在失调。此外，据报道子痫前期孕妇循环中外泌体中 miR-486-5p 增加。然而，miR-486-5p 升高在子痫前期中的作用尚未阐明。
用 miR-486-5p 转染 HTR8/SVneo 滋养细胞，并用定量逆转录聚合酶链反应（RT-qPCR）和 Western blot 检测 ARHGAP5 的表达。通过含有 ARHGAP5 3'-非翻译区（3'UTR）的荧光素酶报告基因检测，确定 ARHGAP5 是否是 miR-486-5p 的直接靶标。通过划痕愈合试验和侵袭试验分别检测迁移和侵袭能力的变化。
根据 RT-qPCR 和 Western blot，miR-486-5p 转染细胞的 ARHGAP5 表达明显降低。双荧光素酶报告基因检测显示 miR-486-5p 直接作用于 ARHGAP5 mRNA 的 3'UTR。miR-486-5p 转染细胞的迁移和侵袭能力受到抑制。用小干扰 RNA 转染下调 ARHGAP5 抑制了滋养细胞的迁移和侵袭，类似于 miR-486-5p 转染。
miR-486-5p 通过抑制 ARHGAP5 的表达至少部分抑制 HTR8/SVneo 细胞的迁移和侵袭能力。这些数据表明 miR-486-5p 参与子痫前期的发病机制，miR-486-5p 是预测子痫前期发病风险的潜在生物标志物。
