Chi Zhenjing, Zhang Muling
Department of Obstetrics, Huai'an First People's Hospital, Huai'an, Jiangsu 223300, P.R. China.
Exp Ther Med. 2018 Dec;16(6):5298-5304. doi: 10.3892/etm.2018.6890. Epub 2018 Oct 23.
Preeclampsia (PE) is the leading cause of maternal and fetal mortality and morbidity. Furthermore, recent studies have reported that miR-145 within the preeclamptic trophoblast debris may cause the high blood pressure via interacting with the maternal endothelium. The aim of the present study was to investigate the functions of miR-145 in PE. Reverse transcription-quantitative polymerase chain reaction and western blotting were used to assess the expression of miR-145 and mucin (MUC1), respectively. TargetScan, miRBase and miRWalk were used to predict the targets of miR-145. Constructed miR-145 mimic plasmids were transfected into HTR-8/SVneo cells for further experiments, including an MTT assay for cell proliferation, Transwell assay for cell invasion and flow cytometry for cell apoptosis analysis. Additionally, the luciferase reporter gene system was employed for target verification. The results demonstrated that miR-145 is downregulated and MUC1 is upregulated in PE tissues and cells compared with normal placenta tissues and cells. The correlation analysis suggests that the expression of miR-145 is negatively correlated with MUC1. Meanwhile, increased proliferation, enhanced invasion and decreased apoptosis of HTR-8/SVneo cells was observed in miR-145 mimic groups compared with mimic control group. Also, the decreased luciferase activity in the miR-145 mimic group indicates that MUC1 may be a target of miR-145. In summary, the results of the present study suggest that miR-145 may serve key roles in the regulation of trophoblast cell proliferation and invasion by targeting MUC1.
子痫前期(PE)是孕产妇和胎儿死亡及发病的主要原因。此外,最近的研究报道,子痫前期滋养层碎片中的miR-145可能通过与母体内皮细胞相互作用导致高血压。本研究的目的是探讨miR-145在子痫前期中的作用。分别采用逆转录定量聚合酶链反应和蛋白质印迹法评估miR-145和黏蛋白(MUC1)的表达。使用TargetScan、miRBase和miRWalk预测miR-145的靶标。将构建的miR-145模拟质粒转染到HTR-8/SVneo细胞中进行进一步实验,包括用于细胞增殖的MTT分析、用于细胞侵袭的Transwell分析和用于细胞凋亡分析的流式细胞术。此外,采用荧光素酶报告基因系统进行靶标验证。结果表明,与正常胎盘组织和细胞相比,子痫前期组织和细胞中miR-145表达下调,MUC1表达上调。相关性分析表明,miR-145的表达与MUC1呈负相关。同时,与模拟对照组相比,miR-145模拟组中HTR-8/SVneo细胞的增殖增加、侵袭增强且凋亡减少。此外,miR-145模拟组中荧光素酶活性降低表明MUC1可能是miR-145的靶标。总之,本研究结果表明,miR-145可能通过靶向MUC1在滋养层细胞增殖和侵袭的调节中发挥关键作用。