miR-18a调节HTR-8细胞中雌激素受体α表达的研究

目的:研究微小RNA-18a(miR-18a)在人正常滋养细胞(HTR8)中对雌激素受体α(ERα)表达的调控作用。方法:miR-18a前体分子miR-18a类似物(miR-18a mimics)、miR-18a空白载体、miR-18a抑制物(miR-18a inhibitor)]转染HTR-8细胞,分为实验组1、阴性对照组(NC组)、实验组2。RT-PCR检测转染后各组中miR-18a的表达情况;RT-PCR与Western-blot检测HTR-8细胞中ERα在mRNA和蛋白水平的表达情况。结果:实验组1细胞中miR-18a的表达与NC组(0.407±0.019)相比明显增高(0.880±0.060),实验组2细胞中miR-18a的表达显著降低(0.160±0.014),差异有统计学意义(P0.05)。转染48小时后实验组1ERαmRNA的表达(0.249±0.003)与NC组(0.313±0.010)相比差异无统计学意义(P0.05);实验组2 ERαmRNA的表达水平明显增高(0.823±0.023),差异有统计学意义(P0.05)。实验组2 ERα蛋白表达水平(1.030±0.006)与NC组(0.960±0.008)相比有增高,实验组1 ERα蛋白表达水平明显降低(0.660±0.013),差异有统计学意义(P0.05)。结论:在人正常滋养细胞中,miR-18a可能在转录和翻译水平均对ERα具有调控作用。

Effect of PE Related miR-18a on Expression of Estrogen Receptor Alpha in HTR-8 Cells

Objective:To study the effect of miR-18a on expression of estrogen receptor alpha in human normal trophoblast cells(HTR-8).Methods:The synthesized miR-18a (miR-18amimics,miR-18a inhibitor and blank carrier) were transfected into HTR-8 cells.Experiment included experimental group 1,pre-miRNA un-transfection group(NC group) and experimental group 3.RT-PCR detected the expression of miR-18a in the three groups.RT-PCR and Western-blot detected the expression of ERα mRNA and protein in the transfected HTR-8 cells.Results:According to the results of RT-PCR,the expressions of miR-18a in experimental group 1 (0.880 ± 0.060) was significantly higher than that of NC group(0.407 ±0.019) (P ＜0.05),and the expressions of miR-18a in experimental group 3(0.160 ±0.014) was significantly lower than that of NC group(0.407 ±0.019) (P＜0.05);After transfected for 48 hours,there was no significant difference between the expression of ERα mRNA between experimental group 1 (0.249 ±0.003) and NC group(0.313 ±0.010) (P＞0.05),the expression of ERα mRNA in transfected cells in experimental group 3 (0.823 ± 0.023)was significantly higher than that of NC group (P ＜ 0.05);Western-blot results showed that the expression of ERα protein of experimental group 3(1.030 ±0.006) was significantly higher than that of NC group(0.960 ±0.008) (P ＜ 0.05);the expression of ERα protein of experimental group 1 (0.660 ±0.013)was significantly lower than that of NC group (0.960 ± 0.008) (P ＜ 0.05).Conclusions:in human normal trophoblast cells,miR-18a may play regulatory role in the expression of ERα both in mRNA and protein levels.