MicroRNA-29a 对人正常滋养细胞迁移和侵袭功能的影响及其可能的机制

Effect of miR-29a on migration and invasion of human normal trophoblast cells and mechanism

摘要:

目的 检测microRNA-29a（miR-29a）在子痫前期孕妇胎盘组织中的表达及其对人正常滋养细 胞系HTR8/Svneo 迁移和侵袭能力的影响及可能的作用机制。方法 收集行剖宫产分娩的30 例子痫前期孕 妇及30 例正常妊娠孕妇的胎盘组织，应用实时聚合酶链反应（real-time PCR）检测胎盘组织miR-29a 的表 达水平。应用生物信息学软件预测miR-29a 的潜在靶基因，选取ITGB 1 作为研究靶基因。将miR-29a 模拟 物（miR-29a mimics）转染人正常滋养细胞系HTR8/Svneo，real-time PCR 检测转染后HTR8/Svneo 细胞中 miR-29a 的表达变化，划痕实验和Transwell 侵袭实验检测转染后HTR8/Svneo 细胞迁移和侵袭能力的变化， Western blot 检测转染后HTR8/Svneo 细胞ITGB1 蛋白的表达变化。结果 real-time PCR 结果显示子痫前 期孕妇胎盘组织中miR-29a 表达水平均较正常妊娠孕妇增高（P <0.05）。人正常滋养细胞系HTR8/Svneo 细 胞转染miR-29a mimics 后，miR-29a 表达水平增高（P <0.05），ITGB1 蛋白表达水平降低（P <0.05），划痕 实验和Transwell 侵袭实验显示细胞迁移和侵袭能力降低（P <0.05）。结论 miR-29a 在子痫前期孕妇胎盘组 织中高表达，可能通过调控ITGB1 的表达参与子痫前期的发生发展。

Abstract:

Objective To explore the expression of miR-29a in placental tissues of patients with preeclampsia and its effect and possible mechanism on invasion and migration of human normal trophoblast cell line HTR8/ Svneo. Methods Placental tissues were collected from 30 pregnant women with preeclampsia and 30 normal pregnant women who delivered by cesarean section. Real-time PCR was used to analyze the expression level of miR- 29a in placental tissues. The potential target genes of miR-29a were predicted by bioinformatics software, and ITGB1 as a target gene was selected. Then, HTR8/Svneo cells were transfected with miR-29a mimics. Real-time PCR was used to explore the expression level of miR-29a and Western blot was used to explore the expression level of ITGB1 protein in HTR8/Svneo cells after transfection. Wound healing assay and Transwell assay were used to analyze the effect of miR-29a on invasion and migration of HTR8/Svneo cells after transfection. Results Real-time PCR showed that the expression of miR-29a in the placental tissues of the patients with preeclampsia was significantly higher than that of normal pregnant women (P < 0.05). After transfection with miR-29a mimics in HTR8/Svneo cells, the expression of miR-29a was significantly increased (P < 0.05), the expression of ITGB1 protein and the capacity of invasion and migration were significantly decreased (P < 0.05). Conclusions The expression of miR-29a is significantly up-regulated in placental tissues of the patients with preeclampsia, and miR-29a may participate in the occurrence and development of preeclampsia by regulating ITGB1 expression.