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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 regulatory effect of microRNA-18a (miR-18a) in preeclampsia on estrogen receptor 1 (ER1) in human normal trophoblast cells (HTR8) and on the cell cycle and apoptosis of HTR8 cells. Methods HTR8 cells were transfected with miR-18a precursor (pre-miR-18a). The experiment was divided into pre-miRNA negative control group, miR-18a overexpression group and miR-18a inhibition group. The expression of miR-18a mRNA in each group was detected by reverse transcription-polymerase chain reaction (RT-PCR). The mRNA and protein levels of miR-18a in each group were detected by RT-PCR and Western blot respectively The changes of cell cycle and apoptosis in each group after transfection were detected by flow cytometry. Results Compared with the pre-miRNA negative control group, miR-18a overexpression and miR-18a inhibition both achieved a significant effect; inhibited miR-18a expression, ER1 mRNA and protein levels increased; overexpression and inhibition of miR-18a on HTR8 cells Cycle had no significant effect; inhibition of miR-18a can increase HTR8 cell apoptosis. Conclusion Inhibition of miR-18a can increase the expression of estrogen receptor 1 in normal human trophoblast cells and promote apoptosis.