Wang Likui, Li Yunguang
Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Wenhua Xi Road, Ji'nan City, Shandong, China.
Arch Med Sci. 2020 Jan 10;18(2):499-522. doi: 10.5114/aoms.2019.91512. eCollection 2022.
This study investigated miR-29b-3p's effects and mechanisms in preeclampsia development.
In this study, we analysed the pathology and expression of miR-29b-3p and B2R mRNA from normal and preeclampsia placenta tissues using hematoxylin and eosin staining and RT-qPCR assay. For cell experiments, we used transwell assay CCK-8, flow cytometry and wound healing assay to determine the effects and correlation of miR-29b-3p and B2R in HTR-8/SVneo cell proliferation, apoptosis, cell cycle, cell invasion and migration in a preeclampsia cell model. Moreover, the mechanisms were determined using Western blot or immunofluorescence in different groups.
Clinical analysis revealed that miR-29b-3p gene expression dramatically increased with increasing degree of preeclampsia ( < 0.001 or < 0.05, respectively). The HTR-8/SVneo cell biological activities of the model group were significantly depressed ( < 0.001). However, with miR-29b-3p inhibitor or B2R transfection, the HTR-8/SVneo cell biological activities significantly recovered ( < 0.001). Western blot assay showed that B2R, VEGF-A, CCND-1, MMP-2 and MMP-9 levels were suppressed in the model group, compared with those in the NC groups ( < 0.001, respectively). With miR-29b-3p inhibitor or B2R transfection, the protein expression levels of B2R, VEGF-A, CCND-1, MMP-2 and MMP-9 dramatically increased ( < 0.001, respectively).
The down-regulation of miR-29b-3p could improve HTR-8/SVneo cell biological activities in a preeclampsia cell model by targeting B2R.
本研究调查了miR-29b-3p在子痫前期发展中的作用及机制。
在本研究中，我们使用苏木精-伊红染色和RT-qPCR检测分析了正常和子痫前期胎盘组织中miR-29b-3p和B2R mRNA的病理及表达情况。对于细胞实验，我们使用Transwell实验、CCK-8、流式细胞术和伤口愈合实验来确定miR-29b-3p和B2R在子痫前期细胞模型中对HTR-8/SVneo细胞增殖、凋亡、细胞周期、细胞侵袭和迁移的影响及相关性。此外，通过蛋白质免疫印迹法或免疫荧光法确定不同组中的机制。
临床分析显示，随着子痫前期程度的增加，miR-29b-3p基因表达显著增加（分别为<0.001或<0.05）。模型组的HTR-8/SVneo细胞生物学活性显著降低（<0.001）。然而，使用miR-29b-3p抑制剂或B2R转染后，HTR-8/SVneo细胞生物学活性显著恢复（<0.001）。蛋白质免疫印迹法检测显示，与NC组相比，模型组中B2R、VEGF-A、CCND-1、MMP-2和MMP-9水平受到抑制（分别为<0.001）。使用miR-29b-3p抑制剂或B2R转染后，B2R、VEGF-A、CCND-1、MMP-2和MMP-9的蛋白质表达水平显著增加（分别为<0.001）。
在子痫前期细胞模型中，miR-29b-3p的下调可通过靶向B2R改善HTR-8/SVneo细胞生物学活性。
