Institute for Fetology, First Hospital of Soochow University, Suzhou 215006, China; Wuxi Maternity and Children Health Hospital Affiliated Nanjing Medical University, Wuxi 214000, China.
Beijing Genomics Institute-Shenzhen, Shenzhen 518083, China.
Biomed Pharmacother. 2017 Oct;94:341-353. doi: 10.1016/j.biopha.2017.07.130. Epub 2017 Aug 1.
Previous studies indicated that microRNAs (miRNAs) were aberrantly expressed in the placentas of patients with Preeclampsia (PE); however, the underlying mechanism still requires further investigation. The aim of this study is to investigate the roles of miR-144 in preeclampsia and the related mechanism. The expression of miR-144 and PTEN in 30 placentas of patients with PE and 30 normal placentas was compared; next, HTR8/SVneo cells were transfected with miR-144 mimics and miR-144 inhibitors and cultured for 48h, and the proliferation and apoptosis, cell migration and invasion of the cells were examined; furthermore, the expression PTEN, Caspase-3 and Bcl-2 was examined; next, dual luciferase reporter assay has been performed to confirm that PTEN is a direct target of miR-144; finally, HTR-8/SVneo cells were transfected with either PTEN overexpression plasmid or PTEN RNAi to determine whether knockdown or overexpression of PTEN can mimic the effect of miR-144 We have observed that the expression of miR-144 was significantly decreased and the expression of PTEN was markedly increased in placentas of patients with PE compared with normal placentas; moreover, transfection of miR-144 mimics in trophoblastic cells induced significant increase in cell proliferation, migration, invasion, and decrease in cell apoptosis, and also affected the cell cycles; on the other hand, transfection of miR-144 inhibitors has shown the opposite effects; furthermore, transient overexpression of miR-144 induced marked decrease in the expression of PTEN, Caspase-3 and increase in expression of Bcl-2 (P<0.01), while transfection of miR-144 inhibitors showed the opposite effects; finally, PTEN has been confirmed as a direct target of miR-144; finally, transfection of PTEN overexpression plasmid or PTEN RNAi can mimic the results of miR-144 inhibitor or miR-144 mimics, respectively. In conclusion, miR-144 was down-regulated in PE, and miR-144 may play important roles in the pathogenesis of PE through targeting PTEN in trophoblastic cells. These results suggested that miR-144 has the potential to become a therapeutic target for the treatment of PE.
先前的研究表明，微 RNA（miRNA）在子痫前期（PE）患者的胎盘中表达异常；然而，其潜在机制仍需要进一步研究。本研究旨在探讨 miR-144 在子痫前期中的作用及其相关机制。比较了 30 例 PE 患者胎盘和 30 例正常胎盘组织中 miR-144 和 PTEN 的表达；接下来，用 miR-144 模拟物和 miR-144 抑制剂转染 HTR8/SVneo 细胞并培养 48h，检测细胞的增殖和凋亡、细胞迁移和侵袭；此外，检测 PTEN、Caspase-3 和 Bcl-2 的表达；接下来，进行双荧光素酶报告基因检测以证实 PTEN 是 miR-144 的直接靶标；最后，用 PTEN 过表达质粒或 PTEN RNAi 转染 HTR-8/SVneo 细胞，以确定 PTEN 的敲低或过表达是否可以模拟 miR-144 的作用。我们观察到，与正常胎盘相比，PE 患者胎盘组织中 miR-144 的表达明显降低，PTEN 的表达明显升高；此外，转染 miR-144 模拟物可诱导滋养细胞显著增加细胞增殖、迁移和侵袭，减少细胞凋亡，并影响细胞周期；另一方面，转染 miR-144 抑制剂则显示相反的效果；此外，瞬时过表达 miR-144 可显著降低 PTEN、Caspase-3 的表达，增加 Bcl-2 的表达（P<0.01），而转染 miR-144 抑制剂则显示相反的效果；最后，证实了 PTEN 是 miR-144 的直接靶标；最后，转染 PTEN 过表达质粒或 PTEN RNAi 可分别模拟 miR-144 抑制剂或 miR-144 模拟物的结果。总之，miR-144 在 PE 中下调，miR-144 可能通过靶向滋养细胞中的 PTEN 在子痫前期的发病机制中发挥重要作用。这些结果表明，miR-144 具有成为治疗 PE 的治疗靶点的潜力。
