【摘要】 目的:研究人类白细胞抗原E(HLA-E)是否参与孕激素对JEG-3细胞VEGF基因表达的调节作用。方法:JEG-3细胞分为5组,接受不同处理:空白对照(组1)、孕激素处理(组2)、转染HLA-E siRNA慢病毒后孕激素处理(组3),HLA-E siRNA慢病毒转染(组4)以及慢病毒阴性对照转染(组5)。48小时后收集细胞,分别采用Real-time PCR和Western blot方法检测JEG-3细胞血管内皮生长因子(VEGF) mRNA及蛋白表达水平。结果:组2 JEG-3细胞的VEGF mRNA和蛋白表达较组1增高,差异有统计学意义(P<0.05);沉默HLA-E基因(组4)导致VEGF mRNA和蛋白表达降低,与组1比较,差异有统计学意义(P<0.05)。组3 JEG-3细胞经HLA-E siRNA慢病毒转染、沉默HLA-E表达后再接受孕激素处理,VEGF mRNA及蛋白表达水平与组1比较差异无统计学意义(P>0.05)。组5与组1比较,VEGF mRNA及蛋白水平比较,差异无统计学意义(P>0.05)。结论:HLA-E介导孕激素对JEG-3滋养细胞VEGF表达的上调,孕激素对JEG-3细胞VEGF的上调作用被HLA-E基因的沉默消除。更多还原

【Abstract】 Objective:To determine if human leukocyte antigen(HLA)-E played a role in the effect of progesterone on the Vascular endothelial growth factor(VEGF) expression in chorion carcinoma JEG-3 cells.Methods:JEG-3 cells were allocated into five groups including group 1(the blank control),group 2(progesterone treatment),group 3(transfection of lentivirus carrying siRNA targeting HLA-E before progesterone treatment),group 4(transfection of lentivirus carrying siRNA targeting HLA-E),and group 5(transfection of the negative control siRNA).The mRNA and protein levels of VEGF in JEG-3 cells were detected by real-time quantitative PCR and Western blot assay,respectively 48 h later.Results:The expression level of VEGF mRNA and protein in JEG-3 was up-regulated after treatment of progesterone(group 2) while down-regulated after transfected with lentivirus carrying siRNA targeting HLA-E(group 4),both with significant differences compared to the control group(P<0.05).The HLA-E in JEG-3 was silenced by transfection of siRNA targeting HLA-E before progesterone treatment,with no difference in VEGF mRNA and protein expression level compared to the control group.Conclusions:HLA-E is involved in the expression level of VEGF and diminishes the up-regulatory effect of progesterone on VEGF in JEG-3.更多还原