目的:通过研究子痫前期胎盘组织中低表达的miR-18b-5p(微小RNA18b-5p)对人滋养细胞系HTR-8迁移能力的影响,进一步探讨miR-18b-5p在子痫前期发病过程中的作用。方法:将化学合成的miR-18b-5p inhibitor、miR-18b-5p inhibitor NC,采用瞬时转染的方法将miR-18b-5p inhibitor转入人滋养细胞系HTR-8中设为实验组;miR-18b-5p inhibitor NC转入HTR-8细胞中设为阴性对照组;空白转染组为空白对照组。应用Realtime RT-PCR技术检测各组miR-18b-5p在mRNA水平的表达,同时采用微孔滤膜培养小室及双室联合培养系统(Transwell实验)检测实验组与对照组细胞迁移能力的变化。结果:Realtime RT-PCR结果显示:转染miR-18b-5p inhibitor后miR-18b-5p的表达量分别与阴性对照组和空白对照组相比明显降低,差异具有统计学意义(P〈0.05)。Transwell实验结果显示miR-18b-5p inhibitor组与两对照组相比细胞的迁移能力明显降低,差异具有统计学意义(P〈0.05)。结论:在HTR-8细胞中降调节miR-18b-5p可以显著的降低细胞的迁移能力,推测病理性低表达的miR-18b-5p有可能通过降低滋养细胞的迁移能力,使妊娠早期细胞滋养细胞从固体绒毛顶端迁出减少,导致覆盖合体滋养细胞进而形成具有增殖能力的细胞簇减少,最终造成滋养层浅表植入,从而引起子痫前期的发生。
Objective: In order to make clear the down-regulated miR-18b-5p plays an important role in the pathogenesis progress of preeclampsia. The effect of miR-18b-5p on the migration capacity of Trophoblast Cell HTR-8 have been studied. Methods: The miR-18b-5p inhibitor and miR-18b-5p inhibitor NC were synthesized by a chemical synthesis process. The technology of transient transfection was used to transfect with miR-18b-5p inhibitor in HTR-8 cells as the experimental group. In the same way, The miR-18b-5p inhibitor NC was transfected into HTR-8 cells as the negative control group. Blank transfection group as the blank control group. The realtime RT-PCR technology have been used to detect each group miR-18b-5p in the expression of mRNA level. At the same time, Transwell experiment were applied to test cell migration ability of experimental group and control group. Results: Compared with control groups, Realtime RT-PCR results showed that the miR-18b-5p amount of expression in miR-18b-5p inhibitor transfection group significantly lower (P 〈0.05 ). Compared with control group, Transwell experiment results showed that the cell migration ability in miR-18b-5P inhibitor group significantly reduced (P〈0.05). Conclusion: MiR-18b-5p can significantly reduce the migration ability of HTR-8 cells, the down-expression of miR-18b -5p can make nourish cells of myometrium from the top of solid fluff out in early pregnancy by reducing the nourish cell migration ability, prompting syncytiotrophoblast that has the proliferation of cell clusters decrease, eventually led to the trophoblast superficial implants, which causing the occurrence of preeclampsia.