siRNA下调AEG-1基因表达对胶质瘤细胞血管拟态形成的影响
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摘要
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-1in 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-1expression on glioma vasculogenic mimicy.Real-time PCR and Western blot were used to detect the mRNA and protein expressions of VEGF and MMP2 in U87cells after treated with AEG-1siRNA,respectively.Immunofluorescence was used to detect the expressions of VEGF and MMP2 in glioma vasculogenic mimicy model in vitro.Results Down-regulation of AEG-1expression 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 downregulation of AEG-1expression 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.
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-1in 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-1expression on glioma vasculogenic mimicy.Real-time PCR and Western blot were used to detect the mRNA and protein expressions of VEGF and MMP2 in U87cells after treated with AEG-1siRNA,respectively.Immunofluorescence was used to detect the expressions of VEGF and MMP2 in glioma vasculogenic mimicy model in vitro.Results Down-regulation of AEG-1expression 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 downregulation of AEG-1expression 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.
引文
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[2]LEE SG,KIM K,KEGELMAN TP,et al.Oncogene AEG-1promotes glioma-induced neurodegeneration by increasing glutamate excitotoxicity[J].Cancer Res,2011,71(20):6514-6523.
[3]STUPP R,MASON WP,VAN DEN BENT MJ,et al.Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma[J].N Engl J Med,2005,352(10):987-996.
[4]KREISL TN,KIM L,MOORE K,et al.Phase II trial of singleagent bevacizumab followed by bevacizumab plus irinotecan at tumor progression in recurrent glioblastoma[J].J Clin Oncol,2009,27(5):740-745.
[5]EL HALLANI S,BOISSELIER B,PEGLION F,et al.A new alternative mechanism in glioblastoma vascularization:Tubular vasculogenic mimicry[J].Brain,2010,133(Pt 4):973-982.
[6]ZHANG S,LI M,GU Y,et al.Thalidomide influences growth and vasculogenic mimicry channel formation in melanoma[J].J Exp Clin Cancer Res,2008,27(1):60.
[7]CONG R,SUN Q,YANG L,et al.Effect of Genistein on vasculogenic mimicry formation by human uveal melanoma cells[J].J Exp Clin Cancer Res,2009,28(1):1-7.
[8]YAO X,PING Y,LIU Y,et al.Vascular endothelial growth factor receptor 2(VEGFR-2)plays a key role in vasculogenic mimicry formation,neovascularization and tumor initiation by glioma stem-like cells[J].PLoS One,2013,8(3):e57188.
[9]ZOU M,ZHU W,WANG L,et al.AEG-1/MTDH-activated autophagy enhances human malignant glioma susceptibility to TGF-beta1-triggered epithelial-mesenchymal transition[J].Oncotarget,2016,7(11):13122-13138.
[10]NOCH E,BOOKLAND M,KHALILI K.Astrocyte-elevated gene-1(AEG-1)induction by hypoxia and glucose deprivation in glioblastoma[J].Cancer Biol Ther,2011,11(1):32-39.
[11]YANG Y,WU J,GUAN H,et al.MiR-136promotes apoptosis of glioma cells by targeting AEG-1and Bcl-2[J].FEBS Lett,2012,586(20):3608-3612.
[12]YU JQ,ZHOU Q,ZHU H,et al.Overexpression of astrocyte elevated gene-1(AEG-1)in cervical cancer and its correlation with angiogenesis[J].Asian Pac J Cancer Prev,2015,16(6):2277-2281.
[13]HUANG LL,WANG Z,CAO CJ,et al.AEG-1associates with metastasis in papillary thyroid cancer through upregulation of MMP2/9[J].Int J Oncol,2017,51(3):812-822.
[14]SHI X,WANG X.The role of MTDH/AEG-1in the progression of cancer[J].Int J Clin Exp Med,2015,8(4):4795-4807.