miR-29b通过调控PTEN的表达对人绒毛膜滋养层细胞生物学行为的影响-现代医学

miR-29b通过调控PTEN的表达对人绒毛膜滋养层细胞生物学行为的影响

单位：株洲市中心医院妇产科, 湖南株洲 412000

分类号：R714

出版年·卷·期（页码）：2021·49·第十期（1200-1207）

摘要：

目的：探讨microRNA-296（miR-29b）及PTEN对人绒毛膜滋养层细胞（HTR-8/SVneo）的增殖、迁移的作用以及两者之间的关系。方法：实时荧光定量聚合酶链反应（qRT-PCR）检测miR-29b和PTEN mRNA的表达；细胞增殖（EDU）检测miR-29b对HTR-8/SVneo细胞增殖能力的情况；Transwell迁移实验检测miR-29b对HTR-8/SVneo细胞迁移能力的影响；TargetScan预测miR-29b与PTEN之间的结合位点；双荧光素酶报告基因检测miR-29b与PTEN之间的相互作用关系；蛋白质免疫印迹（Western blotting）检测miR-29b对PTEN蛋白表达的影响；EDU检测miR-29b靶向PTEN对HTR-8/SVneo细胞增殖能力的情况；Transwell迁移实验检测miR-29b靶向PTEN对HTR-8/SVneo细胞迁移能力的影响。结果：miR-29b在妊娠期糖尿病患者中低表达（P<0.01），PTEN在妊娠期糖尿病患者中高表达（P<0.01）；过表达miR-29b使HTR-8/SVneo细胞中EDU细胞阳性比明显降低（P<0.01），迁移细胞数目明显减少（P<0.01）；而沉默miR-29b使HTR-8/SVneo细胞中EDU细胞阳性比明显增加（P<0.01），迁移细胞数目增加（P<0.001）；miR-29b与PTEN存在靶向关系，且两者具有相互抑制的作用；过表达PTEN可逆转miR-29b过表达引起的HTR-8/SVneo细胞的增殖能力和迁移能力下降（P<0.01）；沉默PTEN可逆转miR-29b低表达引起的HTR-8/SVneo细胞的增殖能力和迁移能力增加（P<0.01）。结论：miR-29b通过调控PTEN的表达调节滋养细胞的增殖能力和迁移能力。

Objective: To explore the effect of microRNA-29b(miR-29b) and PTEN on the proliferation and migration of human trophoblasts (HTR-8/SVneo) and the relationship. Methods: qRT-PCR was used to detect the expression of miR-29b and PTEN mRNA; EDU was used to detect the effect of miR-29b on the proliferation of HTR-8/SVneo cells; Transwell migration test was used to detect the effect of miR-29b on the migration ability of HTR-8/SVneo cells; TargetScan was used to predict the binding site between miR-29b and PTEN; dual luciferase reporter gene was used to detect the interaction between miR-29b and PTEN; Western blottingwas used to detect the effect of miR-29b on PTEN protein expression; EDU was used to detect miR-29b targeting PTEN on the proliferation of HTR-8/SVneo cells; Transwell migration test was used to examined the effect of miR-29b targeting PTEN on the migration ability of HTR-8/SVneo cells. Results: miR-29b waslower expressed in gestational diabetes patients(P<0.01), PTEN was highly expressed in gestational diabetes patients(P<0.01); Overexpression of miR-29b significantly reduced the positive ratio of EDU cells in HTR-8/SVneo cells (P<0.01), and significantly reduced the number of migrated cells (P<0.01); while silencing miR-29b significantly increased the positive ratio of EDU cells in HTR-8/SVneo cells, and increased the number of migrated cells (P<0.001); miR-29b had a targeting relationship with PTEN, and both have mutual inhibition effects; Overexpression of PTEN can reverse the reduction of proliferation and migration ability of HTR-8/SVneo cells caused by miR-29b overexpression (P<0.01); Silencing PTEN can reverse the increased proliferation and migration ability of HTR-8/SVneo cells caused by low expression of miR-29b (P<0.01). Conclusion: miR-29b regulates the proliferation and migration of trophoblast cells by regulating the expression of PTEN.

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