Zheng Dongying, Hou Yue, Li Yuanyuan, Bian Yue, Khan Muhanmmad, Li Fan, Huang Ling, Qiao Chong
Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shenyang, China.
Department of Obstetrics and Gynecology, Second Affiliated Hospital of Dalian Medical University, Dalian, China.
Front Genet. 2020 Mar 3;11:188. doi: 10.3389/fgene.2020.00188. eCollection 2020.
Preeclampsia is a lethal pregnancy specific hypertensive disorder involving multisystem. Despite extensive studies to investigate the causes of preeclampsia, the pathogenesis still remains largely unknown. Long non-coding RNAs (lncRNAs) are a diverse class of non-translated RNAs which play a crucial part in various biological phenomena. Although lncRNA Growth Arrest-Specific 5 (GAS5) aberrantly expressed in multiple cancer tissues and is implicated in multiple biological processes of tumor cells, little is known about its role in preeclampsia. In this study, 40 patients with preeclampsia and 32 gestational age matched normotension pregnant women were recruited. Using quantitative real-time polymerase chain reaction (qRT-PCR), we found higher expression of GAS5 in placenta of preclamsia affected women. The level of GAS5 existed strongly in correlation with Thrombin Time indicating coagulation function and other clinical parameters by Pearson correlation analysis. Then we constructed the GAS5 lentivirus expression vectors and transfected into human trophoblast cell lines HTR-8/SVneo and JEG-3. Using cell culture studies, we found an inhibited effect of GAS5 on proliferative ability, migratory ability and invasive ability however; no effect on apoptosis was detected. Further mechanistic analysis found that GAS5 modulated microRNA-21 (miR-21) in an opposite variation tendency by qRT-PCR and rescue experiment. In addition, inhibition of GAS5 promoted the activation of PI3K/AKT signaling pathway and its downstream proteins covering MMP-9 and TP53 as evident from our qRT-PCR and western blot analyses. Thus, we suggested that GAS5 might involve in pregnancy with preeclampsia by influencing the biological functions of trophoblast cells through the regulation of miR-21 and activation of PI3K/AKT signaling pathway and its downstream targets, which may contribute to reveal the nature of preeclampsia.
子痫前期是一种致命的妊娠特异性多系统高血压疾病。尽管对其病因进行了广泛研究，但发病机制仍不清楚。长链非编码RNA（lncRNAs）是一类多样的非翻译RNA，在各种生物现象中起关键作用。虽然lncRNA生长停滞特异性5（GAS5）在多种癌组织中异常表达并参与肿瘤细胞的多种生物学过程，但其在子痫前期中的作用知之甚少。本研究招募了40例子痫前期患者和32例孕周匹配的血压正常孕妇。通过定量实时聚合酶链反应（qRT-PCR），我们发现子痫前期患者胎盘组织中GAS5表达较高。Pearson相关性分析显示，GAS5水平与凝血酶时间（反映凝血功能）及其他临床参数密切相关。随后我们构建了GAS5慢病毒表达载体并转染到人滋养层细胞系HTR-8/SVneo和JEG-3中。通过细胞培养研究，我们发现GAS5对细胞增殖能力、迁移能力和侵袭能力有抑制作用，但未检测到对细胞凋亡的影响。进一步的机制分析发现，通过qRT-PCR和拯救实验，GAS5与微小RNA-21（miR-21）呈相反的变化趋势。此外，qRT-PCR和蛋白质印迹分析表明，抑制GAS5可促进PI3K/AKT信号通路及其下游蛋白（包括MMP-9和TP53）的激活。因此，我们认为GAS5可能通过调节miR-21以及激活PI3K/AKT信号通路及其下游靶点来影响滋养层细胞的生物学功能，从而参与子痫前期的发生，这可能有助于揭示子痫前期的本质。
