siRNA下调AEG-1基因表达对胶质瘤细胞血管拟态形成的影响

Effects of down-regulation of astrocyte elevated gene-1 expression by siRNA on glioma vasculogenic mimicy

目的探讨下调星型细胞上调基因-1(AEG-1)表达对胶质瘤细胞血管拟态(VM)形成的影响及其可能的机制。方法利用siRNA下调U87胶质瘤细胞中AEG-1表达,通过体外构建胶质瘤VM模型,研究下调AEG-1表达对胶质瘤VM形成的影响;Real-time PCR及Western blot分别检测AEG-1表达下调对U87细胞中血管内皮生长因子(VEGF)、基质金属蛋白酶-2(MMP2)mRNA及蛋白表达的影响;免疫荧光法检测AEG-1表达下调对VM模型中VEGF及MMP2表达的影响。结果 siRNA下调AEG-1表达能显著降低U87细胞体外VM的形成(P<0.01),同时下调AEG-1基因能够显著抑制U87细胞及其构成的VM中VEGF及MMP2的表达(P<0.01)。结论 siRNA下调AEG-1表达能显著抑制胶质瘤细胞VM的形成,其部分机制可能是通过下调VEGF及MMP2表达实现。

Objective To investigate the effects of down-regulation of astrocyte elevated gene-1(AEG-1)expression by siRNA on glioma vasculogenic mimicy in vitro.Methods The expression of AEG-1 in U87 glioma cells was down-regulated by siRNA.An in vitro model of glioma vasculogenic mimicy was established to evaluate the effect of down-regulation of AEG-1 expression on glioma vasculogenic mimicy.Real-time PCR and Western blot were used to detect the mRNA and protein expressions of VEGF and MMP2 in U87 cells after treated with AEG-1 siRNA,respectively.Immunofluorescence was used to detect the expressions of VEGF and MMP2 in glioma vasculogenic mimicy model in vitro.Results Down-regulation of AEG-1 expression by siRNA could significantly inhibit the glioma vasculogenic mimicy in vitro(P<0.01).Meanwhile,It could also inhibit the expressions of VEGF and MMP2 in glioma cells and glioma vasculogenic mimicy in vitro(P<0.01).Conclusion These results suggest that down-regulation of AEG-1 expression by siRNA can significantly inhibit the glioma vasculogenic mimicy in vitro,and these effects may be realized partly through regulation of VEGF and MMP2 expression.