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微小RNA-29b-3p通过调控缓激肽B2受体影响人绒毛外滋养层细胞的生长和生物学功能。

MiR-29b-3p affects growth and biological functions of human extravillous trophoblast cells by regulating bradykinin B2 receptor.

作者信息

Wang Likui, Li Yunguang

机构信息

Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Wenhua Xi Road, Ji'nan City, Shandong, China.

出版信息

Arch Med Sci. 2020 Jan 10;18(2):499-522. doi: 10.5114/aoms.2019.91512. eCollection 2022.

Abstract

INTRODUCTION

This study investigated miR-29b-3p's effects and mechanisms in preeclampsia development.

MATERIAL AND METHODS

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.

RESULTS

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).

CONCLUSIONS

The down-regulation of miR-29b-3p could improve HTR-8/SVneo cell biological activities in a preeclampsia cell model by targeting B2R.

摘要

引言

本研究调查了miR-29b-3p在子痫前期发展中的作用及机制。

材料与方法

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。

结果

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)。

结论

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/979f/8924841/059af60417c0/AMS-18-2-109125-g001.jpg

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