VE-statin/Egf17-siRNA抑制恶性胶质瘤诱导内皮细胞血管生成能力的体外研究

Inhibitory effect of VE-statin/Egfl7-siRNA on angiogenesis of endothelial cells induced by malignant glioma in vitro

目的通过细胞培养及RNA干扰技术探讨类表皮生长因子域7(VE-statin/Egfl 7)在恶性胶质瘤血管生成中的作用和机制。方法Transwell培养技术建立U 251-HUVEC共培养系统,体外模拟恶性胶质瘤与内皮细胞的相互作用;并通过构建靶向VE-statin/Egfl7基因的siRNA慢病毒载体抑制U251和HUVEC细胞中该基因的表达。通过内皮细胞增殖、粘附、迁移及成管实验检测该基因在体外恶性胶质瘤微环境中对血管生成的影响。结果沉默VE-statin/Egfl 7基因表达后,HUVEC生长出现暂时性减缓,但很快恢复正常增殖状态,而且内皮细胞的迁移能力不受影响,但是内皮细胞的粘附能力明显受到抑制;成管实验发现,VE-statin/Egfl7基因沉默后内皮细胞不能形成毛细血管样结构。结论 VE-statin/Egfl7可通过调节内皮细胞的粘附性而在恶性胶质瘤血管生成过程的血管管腔形成过程中起着关键性调控作用。

Objective To investigate the function and mechanism of epidermal growth factor-like domain 7 （VE-statin/Egf17） in malignant glioma angiogenesis by cell culture and RNA interference. Methods Interaction between tumor cells and endothelial cells in malignant glioma was simulated by transwell-based co-culture of U251 and HUVEC cells. A lentivirus system carrying siRNA was used to inhibit the expression of VE-statin/Egf17 in HUVEC cells and U251 cells. The effects of VE-statin/Egf17 on angiogenesis in malignant glioma microenvironment were evaluated by endothelial cell proliferation, adhesion, migration, and tube formation assay. Results After VE-statin/Egf17 gene silencing, a temporary growth inhibition was observed in HUVEC cells, followed by immediate recovery of normal proliferation. The endothelial cell migration was not affected, while the adhesion of endothelial ceils was significantly inhibi- ted. VE-statin/Egf17 gene silencing also significantly suppressed the formation of capillary-like structures by endothelial cells. Conclu- sions VE-statin/Egf17 plays a key role in the formation of vascular tubular structure in malignant glioma angiogenesis via regulation of endothelial cell adhesion.