MiR-26b在子痫前期胎盘中差异表达的临床意义

Micro RNA-26b affects proliferation of placenta trophoblast cells in preeclampsia by targeting smad1

目的探讨子痫前期胎盘组织中miR-26b差异表达及其临床意义。方法收集子痫患者与健康人胎盘组织各30例,RT-qPCR和Western blotting检测组织中miR-26b与Smad1的表达;转染miR-26b的mimics到HTR-8/SVneo细胞中,MTT法比较转染组、NC对照组细胞增殖情况,流式细胞术检测各组细胞周期情况;同时RT-qPCR、Western blotting检测各组miR-26b和Smad1的表达情况,并运用双荧光素酶报告检测试验验证miR-26b与靶标Smad1之间靶向关系。结果 miR-26b在子痫前期胎盘组织中高表达,且Smad1的表达与miR-26b的表达相反;滋养层细胞中miR-26b过表达会抑制细胞增殖(P<0.01),并将细胞阻滞在G0/G1期。miR-26b过表达会降低Smad1 mRNA与蛋白的表达(P<0.01)。双荧光素酶报告基因实验证明Smad1是miR-26b的直接靶点。结论 miR-26b可能通过靶向Smad1调控HTR-8/SVneo细胞的增殖与周期。

Objective To investigate the targeting effect and potential mechanism of miR-26b in preeclampsia. Methods 30 cases of preeclampsia patients and 30 cases of normal pregnant women were chosen and trophoblast cells were taken from their placenta tissue. The cell line HTR-8/SVneo cells were transfected with miR-26b mimics and the efficiency of transfection was evaluated by RT-qPCR. MTT proliferation assay was conducted to compare the proliferation in miR-26bmimics transfected and NC groups. The cell cycle of each group were determined by the flow cytometry .The expression of miR-26b and Smadl were determined by RT-qPCR and western blotting assays. And Dual-Luciferase reporter assay was performed to identify the binding site of Smadl and miR-26b. Results MicroRNA-26b were up-regulated expressed in the placenta tissue of preeclampsia patients, with Smadl down-regulated expressed. Transient over-expression of miR-26b in HTR-8/SVneo cells induced significant decrease in cell proliferation （P〈0.01） and blocked the cells in G0/G1 phase, and also reduce the expression of Smadl on both mRNA and protein level. Moreover, dual-luciferase reporter assay confirmed that Smadl is a direct target of miR-26b. Conclusion MiR-26b may regulate the proliferation of HTR-8/SVneo cells via targeting Smadl.