沉默星形细胞上调基因-1联合全反式维甲酸抑制胶质瘤血管拟态形成的机制分析  

Effects of downregulation of astrocyte up-regulated gene-1 expression combined with all-trans retinoic acid on vascular mimicry in glioma

作　　者：赵婉莹 张斌斌[2] 李瑞春[2] 梁晨 Zhao Wanying;Zhang Binbin;Li Ruichun;Liang Chen(International Medical Center,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an,Shaanxi 710061,China;Department of Neurosurgery,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an,Shaanxi 710061,China)

机构地区：[1]西安交通大学第一附属医院涉外病房,陕西西安710061 [2]西安交通大学第一附属医院神经外科,陕西西安710061

出　　处：《中国微侵袭神经外科杂志》2024年第1期24-29,共6页Chinese Journal of Minimally Invasive Neurosurgery

基　　金：陕西省重点研发计划项目(编号:2021SF-298)。

摘　　要：目的研究沉默星形细胞上调基因-1(AEG-1)联合全反式维甲酸(ATRA)对胶质瘤血管拟态(VM)形成的影响及其可能的机制。方法构建AEG-1 shRNA慢病毒稳染U87胶质瘤细胞并用含20μmol/L全反式维甲酸(ATRA)的培养基干预,分组:空白对照组(U87细胞)、ATRA组(U87细胞+20μmol/L ATRA),siCon组(U87-siCon细胞)、siCon+ATRA组(U87-siCon细胞+20μmol/L ATRA),siAEG-1组(U87-siAEG-1细胞)和siAEG-1+ATRA组(U87-siAEG-1细胞+20μmol/L ATRA)。体外管腔形成实验评估U87细胞体外血管拟态形成能力,实时PCR及Western-Blot检测U87细胞中血管拟态形成相关基因表达情况。建立裸鼠胶质瘤皮下移植瘤模型并以10mg/kg剂量给予腹腔注射ATRA干预,分为:对照组(U87细胞造模)、ATRA组(U87细胞造模+ATRA干预),siCon组(U87-siCon细胞造模)、siCon+ATRA组(U87-siCon细胞造模+ATRA干预),siAEG-1组(U87-siAEG-1细胞造模)与siAEG-1+ATRA组(U87-siAEG-1细胞造模+ATRA干预)。定期测量皮下移植瘤大小并绘制肿瘤生长曲线,CD34-PAS双染检测移植瘤中血管拟态。结果沉默AEG-1联合ATRA干预(即siAEG-1+ATRA组)显著抑制胶质瘤细胞及胶质瘤模型中血管拟态形成及皮下移植瘤生长,干预后胶质瘤细胞中基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9),钙黏蛋白(VE-cadherin)表达明显下调。上述结果与对照组比较,差异均具有统计学意义(均P<0.05)。结论沉默AEG-1联合ATRA能显著抑制胶质瘤血管拟态形成及肿瘤生长,其机制可能与抑制MMP-2、MMP-9以及VE-cadherin表达有关。Objective To investigate the effects and potential mechanisms of downregulation of astrocyte up-regulated gene-1(AUG-1)expression combined with all-trans retinoic acid(ATRA)on vasculogenic mimicry(VM)formation in glioma.Methods U87 glioma cells were transfected with AEG-1 shRNA lentiviral vectors(U87-siAEG-1)and incubated in a medium containing 20μmol/L ATRA.A Matrigel-based tube formation assay was performed to evaluate VM formation in vitro.They were divided into control group(U87 cells),ATRA group(U87 cells+20μmol/L ATRA),siCon group(U87-siCon cells),siCon+ATRA group(U87-siCon cells+20μmol/L ATRA),siAEG-1 group(U87-siAEG-1 cells)and siAEG-1+ATRA group(U87-siAEG-1 cells+20μmol/L ATRA).In vitro lumen formation assay was performed for evaluating in vitro vascular mimicry formation ability of U87 cells.Realtime PCR and Western-blot analysis were conducted to investigate the expression of related gene.Glioma xenograft models were generated via subcutaneous implantation of glioma cells in nude mice.Tumor-bearing mice received an intraperitoneal injection of 10mg/kg ATRA.It was divided into:control group(U87 cell modeling),ATRA group(U87 cell modeling+ATRA intervention),siCon group(U87-siCon cell modeling),siCon+ATRA group(U87-siCon cell modeling+ATRA intervention),siAEG-1 group(U87-siAEG-1 cell modeling)and siAEG-1+ATRA group(U87-siAEG-1 cell modeling+ATRA intervention).CD34/periodic acid-Schiff double staining was performed to detect VM channels in vivo.The size of tumors was measured,and the tumor growth curve was drawn to evaluate the tumor growth.Results A combination of ATRA intervention and downregulation of AEG-1 expression(siAEG-1+ATRA group)significantly decreased VM formation of glioma in vitro and in vivo,and significantly inhibit the growth of subcutaneous glioma xenografts.The expression levels of matrix metalloproteinase(MMP)-2,MMP-9 and VE-cadherin in glioma cells significantly decreased after intervention.Compared with the control group,the difference was statistically significant(all P<0.05).Con

关 键 词：神经胶质瘤 星形细胞上调基因-1 全反式维甲酸 血管拟态 

分 类 号：R730.264[医药卫生—肿瘤]