摘要目的:探讨微小RNA（miR）-216a-5p在妊娠期糖尿病（GDM）患者胎盘中的表达及其对滋养层HTR-8/SVneo细胞侵袭、迁移的影响及可能机制。方法:收集2018年10月至2019年10月烟台市烟台山医院的28例GDM孕妇（GDM组）和28例正常孕妇（对照组）胎盘组织，将体外培养HTR-8/SVneo细胞分为空白组（正常培养）、模拟物（mimics）-NC组（转染mimics-NC）和miR-216a-5p mimics组（转染miR-216a-5p mimics），采用实时荧光定量PCR检测胎盘组织和各组HTR-8/SVneo细胞中miR-216a-5p表达水平，Transwell实验检测各组HTR-8/SVneo细胞侵袭和迁移，免疫印迹法（Western blot）检测各组HTR-8/SVneo细胞中E-钙黏附蛋白（E-cadherin）、波形蛋白（Vimentin）、锌指E盒结合蛋白-1（ZEB1）和N-钙黏附分子（N-cadherin）蛋白表达，双荧光素酶报告基因实验检测miR-216a-5p和ZEB1的靶向关系。结果:与对照组比较，miR-216a-5p在GDM组胎盘组织中表达明显升高（2.86±0.35 vs 0.96±0.07, P<0.05）；与mimics-NC组比较，miR-216a-5p mimics组细胞中miR-216a-5p和E-cadherin蛋白表达水平明显升高[（1.01±0.08）vs（22.48±3.26），（0.25±0.03）vs（0.68±0.05）]，而细胞侵袭、迁移能力和细胞中Vimentin、N-cadherin、ZEB1蛋白表达水平明显降低[（106.83±9.35）vs（59.16±5.28），（205.48±18.25）vs（87.32±6.50），（0.65±0.05）vs（0.35±0.02），（0.55±0.04）vs（0.32±0.03），（0.50±0.03）vs（0.36±0.02）]（ P<0.05）。双荧光素酶报告基因实验结果证实miR-216a-5p可与ZEB1靶向结合。 结论:miR-216a-5p在GDM患者胎盘组织中表达上调，可能通过靶向调控ZEB1表达抑制滋养层HTR-8/SVneo细胞转移。

abstractsObjective:To investigate the expression of microRNA (miR) -216a-5p in placenta of gestational diabetes mellitus (GDM) patients, its effects on invasion and migration of trophoblast HTR-8/SVneo cells and possible mechanisms.Methods:Placenta tissues of 28 pregnant women with GDM and 28 normal pregnant women (control group) were collected, HTR-8/SVneo cells were divided into three groups: blank group (normal culture) , mimics-NC group (transfection of mimics-NC) and miR-216a-5p mimics group (transfection of miR-216a-5p mimics) , the expression level of miR-216a-5p in placenta and HTR-8/SVneo cells was detected by real-time fluorescence quantitative PCR, the invasion and migration of HTR-8/SVneo cells were detected by Transwell test, Western blot was used to detect the protein expressions of E-cadherin, Vimentin, zinc finger E-box binding protein-1 (ZEB1) and N-cadherin in HTR-8/SVneo cells, and the targeting relationship between miR-216a-5p and ZEB1 was detected by double luciferase reporter gene assay.Results:Compared with those in the control group, the expression of miR-216a-5p was significantly higher in placental tissue of GDM group ( P<0.05) ; the protein expression levels of miR-216a-5p and E-cadherin in miR-216a-5p mimics group was significantly higher than those in mic-nc group, however, the invasion, migration and expression levels of Vimentin, N-cadherin and ZEB1 were significantly lower ( P<0.05) . In addition, the results of double luciferase reporter gene showed that miR-216a-5p could bind to ZEB1 targetingly. Conclusions:MiR-216a-5p is up-regulated in placental tissues of GDM patients, which may inhibit the transfer of trophoblast HTR-8/SVneo cells by targetingingly regulating the expression of ZEB1.