VASH2对脑胶质瘤血管新生的影响机制

Role of VASH2 in regulation of angiogenesis in glioma

摘要:

目的 通过观察血管抑制素-2(VASH2)对脑胶质瘤微血管内皮细胞增殖、迁移和管形成的调控作用,探究其对脑胶质瘤血管新生的影响机制。方法 构建人脑胶质瘤微血管内皮细胞模型,通过稳定转染获得VASH2过表达和表达沉默的细胞系,在后续实验中分组,分别为空白对照组、VASH2过表达阴性对照空质粒组[VASH2(+)NC 组]、VASH2过表达组[VASH2(+)组]、VASH2表达沉默阴性对照空质粒组[VASH2 (-) NC 组]和VASH2表达沉默组[VASH2(-)组]。通过细胞生长抑制实验检测细胞增殖能力的变化；体外管形成实验检测细胞管形成能力的变化；细胞迁移实验检测细胞迁移能力的变化。应用 GraphPad Prism v5.01统计软件进行数据分析。多组间比较采用单因素方差分析,组间两两比较采用Newman-Keuls分析。结果 在人脑胶质瘤微血管内皮细胞中上调VASH2的表达,VASH2过表达组与 VASH2过表达阴性对照空质粒组相比较,细胞的增殖能力、迁移能力和管形成能力均明显增强(P<0.05) ,表明VASH2过表达能够促进胶质瘤血管的新生。在人脑胶质瘤微血管内皮细胞中下调VASH2的表达,VASH2表达沉默组与VASH2表达沉默阴性对照空质粒组相比较,细胞的增殖能力、迁移能力和管形成能力均明显减弱(P<0.05),表明VASH2沉默能够抑制胶质瘤血管的新生。结论 VASH2在胶质瘤管形成的调控方面扮演了重要的角色,可能为胶质瘤的抗血管治疗提供新策略。

Abstract:

Objective To explore the mechanism of vasohibin 2 (VASH2) on angiogenesis of glioma by regulating the proliferation, migration and tube formation of glioma microvascular endothelial cells. Methods After cell model of human glioma microvascular endothelial cells (GECs) was obtained, VASH2 overexpression and silencing cell lines were established by stable transfection. In the follow-up experiment, they were divided into control group, VASH2 overexpression negative control empty plasmid[VASH2(+)NC] group, VASH2 overexpression [ VASH2(+) ]group, VASH2 expression silencing negative control empty plasmid [VASH2 (-) NC ]group and VASH2 expression silencing [VASH2(-)] group. Cell counting kit-8 (CCK-8) assay was used to detect cell proliferation, tube formation assay was employed to observe the changes of cell tube formation, and transwell assay was adopted to measure cell migration ability. GraphPad Prism v5.01 statistical software was used for data analysis. One-way ANOVA was employed for comparison between multiple groups and Newman-Keuls analysis was used for pairwise comparison between groups.Results When the expression of VASH2 was up-regulated in GECs, there was significant difference between VASH2(+) group and VASH2(+)NC group (P<0.05). The cell proliferation ability, migration ability and tube formation ability were significantly enhanced. It showed that VASH2 overexpression can promote the angiogenesis of glioma. When the expression of VASH2 was down-regulated, there was significant difference between VASH2(-) group and VASH2 (-) NC group (P<0.05). The proliferation ability, migration ability and tube formation ability of cells were significantly weakened, indicating that VASH2 silencing can inhibit the angiogenesis of glioma. Conclusion VASH2 plays an important role in the regulation of glioma angiogenesis. VASH2 may be considered as a potential target in the therapy for glioma in the future.