摘要
目的研究子痫前期相关微小RNA-18a(miR-18a)对人正常滋养细胞(HTR8)中雌激素受体1(ER1)的调控作用及对HTR8细胞周期及凋亡的影响。方法使用miR-18a前体分子(pre-miR-18a)分组转染HTR8细胞,实验分为pre-miRNA阴性对照组、miR-18a过表达组、miR-18a抑制组。反转录PCR(RT-PCR)检测转染后各组细胞中miR-18a mRNA的表达水平;RT-PCR及Western blot法分别检测各组细胞中miR-18a的靶基因ER1在mRNA和蛋白水平的表达;流式细胞术检测转染后各组细胞周期及凋亡的变化。结果与pre-miRNA阴性对照组相比,miR-18a过表达和miR-18a抑制均获得明显效果;抑制miR-18a表达后,ER1 mRNA和蛋白水平增加;过表达和抑制miR-18a对HTR8细胞周期无明显影响;抑制miR-18a可增加HTR8细胞凋亡。结论抑制miR-18a的水平增加HTR8人正常滋养细胞雌激素受体1的表达并促进细胞凋亡。
Objective To investigate the role of pre-eclampsia related miR-18 a in regulating estrogen receptor 1（ ER1）expression,cell cycle and apoptosis in HTR8 human trophoblasts. Methods HTR8 cells were transfected with synthesized negative control（ NC） miRNA,miR-18 a and miR-18 a inhibitor,respectively. The mRNA expressions of miR-18 a and ER1 were determined by reverse transcription PCR（ RT-PCR）. The protein expression level of ER1 was examined by Western blotting. The cell cycle and apoptosis were evaluated by flow cytometry. Results The mRNA level of miR-18 a was significantly up-regulated in miR-18a-transfected group,while down-regulated in miR-18 a inhibitor group as compared with the NC group in HTR8 cells. Compared with the NC group,both mRNA and protein expression levels of ER1 were significantly higher in the miR-18 a inhibitor group. Cell cycle analysis showed that no significant difference was observed among the three groups. The apoptosis rate was significantly higher in the miR-18 a inhibitor group as compared with the other two groups.Conclusion Inhibition of miR-18 a increases the expression of ER1 and promotes apoptosis in HTR8 human trophoblasts.
作者
杨洋
张少华
李亚军
韩彬
马媛
YANG Yang ZHANG Shaohua LI Yajun HAN Bin MA Yuan(First Affiliated Hospital,Xi＇an Medical University,Reproductive Medicine Center,Tangdu Hospital,Fourth Military Medical Universit)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2017年第8期1102-1107,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
陕西省科技计划项目(S2016YFSF0367)
