Reproductive Medical Center, The First Affiliated Hospital of Guangxi Medical University, Nanning, China; Departments of Gynecology and Obstetrics, The First Affiliated Hospital of Xinxiang Medical University, Weihui, China.
Department of Obstetrics, The First Affiliated Hospital of Guangxi Medical University, Nanning, China.
Life Sci. 2018 Sep 15;209:132-139. doi: 10.1016/j.lfs.2018.08.005. Epub 2018 Aug 3.
Deficiency in trophoblast invasion is causally linked to the pathogenesis of preeclampsia. Tumor necrosis factor-alpha (TNF-α) shows the ability to suppress the invasiveness of trophoblasts, while cysteine-rich 61 (Cyr61) exerts an opposite function in trophoblast invasion. This study was designed to check the hypothesis that cysteine-rich 61 (Cyr61) may be involved in the anti-invasive activity of TNF-α in trophoblasts. To this end, we examined the effect of TNF-α treatment on Cyr61 expression in HTR-8/SVneo trophoblast cells and investigated the mechanism for the regulation of Cyr61 by TNF-α. Gain-of-function experiments were performed to clarify the role of Cyr61 in TNF-α-dependent suppression of trophoblast invasion. It was found that TNF-α at 1 and 10 ng/mL reduced Cyr61 protein levels by 30 and 80%, respectively, in HTR-8/SVneo cells, but did not affect the mRNA expression of Cyr61. Mechanistically, microRNA (miR)-145-5p was stimulated by TNF-α and negatively regulated the expression of Cyr61 via interaction with its 3'-untranslated region. Functionally, overexpression of miR-145-5p significantly impaired the migration and invasion of HTR-8/SVneo cells. Depletion of miR-145-5p rescued HTR-8/SVneo cells from TNF-α-mediated invasion suppression, which coincided with prevention of Cyr61 downregulation by TNF-α. In addition, overexpression of Cyr61 partially restored the invasion of HTR8/SVneo cells co-transfected with miR-145-5p mimic or exposed to TNF-α. Taken together, miR-145-5p-mediated downregulation of Cyr61 is required for the anti-invasive effect of TNF-α on trophoblasts.
滋养细胞侵袭缺陷与子痫前期的发病机制有关。肿瘤坏死因子-α（TNF-α）具有抑制滋养细胞侵袭的能力，而富含半胱氨酸的 61（Cyr61）在滋养细胞侵袭中发挥相反的作用。本研究旨在验证富含半胱氨酸的 61（Cyr61）可能参与 TNF-α在滋养细胞中的抗侵袭活性的假说。为此，我们研究了 TNF-α处理对 HTR-8/SVneo 滋养细胞中 Cyr61 表达的影响，并探讨了 TNF-α调节 Cyr61 的机制。通过功能获得实验阐明了 Cyr61 在 TNF-α依赖性抑制滋养细胞侵袭中的作用。结果发现，1 和 10ng/mL 的 TNF-α分别使 HTR-8/SVneo 细胞中的 Cyr61 蛋白水平降低了 30%和 80%，但不影响 Cyr61 的 mRNA 表达。机制上，TNF-α刺激 microRNA（miR）-145-5p，通过与 3'-非翻译区的相互作用负调控 Cyr61 的表达。功能上，miR-145-5p 的过表达显著损害了 HTR-8/SVneo 细胞的迁移和侵袭。miR-145-5p 的耗竭挽救了 HTR-8/SVneo 细胞免受 TNF-α介导的侵袭抑制，同时预防了 Cyr61 被 TNF-α下调。此外，Cyr61 的过表达部分恢复了共转染 miR-145-5p 模拟物或暴露于 TNF-α的 HTR8/SVneo 细胞的侵袭。综上所述，miR-145-5p 介导的 Cyr61 下调是 TNF-α对滋养细胞的抗侵袭作用所必需的。
