目的:探讨肿瘤血管内皮细胞(GVEC)在胶质瘤肿瘤细胞迁移过程中的作用。方法:分别培养正常血管内皮细胞(VEC)及胶质瘤GVEC,在体外与胶质瘤细胞株(CGH5)共培养,并以单独培养的CHG5细胞作为对照。通过Transwell培养板迁移实验分析GVEC对CHG5细胞迁移能力的影响;免疫荧光染色观察GVEC对CHG5细胞骨架的影响;Western blotting检测共培养后CHG5细胞Rac1和Cdc42表达的变化。结果:CHG5与GVEC共培养后迁移能力显著高于与VEC共培养和单独培养的CHG5细胞(P<0.05)。细胞骨架免疫荧光染色显示,与GVEC共培养的CHG5细胞的微管蛋白形态发生改变,细胞核附近微管蛋白较粗厚,呈圆环状,放射状排列。Western blotting结果表明,与GVEC共培养的CHG5细胞Rac1和Cdc42表达明显增高(P<0.05),但与VEC共培养的CHG5细胞表达无明显变化(P>0.05)。结论:肿瘤细胞血管在胶质瘤细胞迁移的过程中至关重要,GVEC可以通过改变细胞微管蛋白结构,并上调Rac1和Cdc42蛋白的表达,促进胶质瘤细胞的迁移运动。
Objective: To investigate the role of tumor vascular endothelial cells(GVEC) in the migration of glioma tumor cells. Methods: The vascular endothelial cells(VEC) and GVEC were isolated,and co-cultured with glioma cell line CGH5 in vitro ,respectively.The CHG5 cells were set as the control.The effects of GVEC on the migration ability and cytoskeleton of CHG5 cells were analyzed using the Transwell migration assay and immunofluorescence staining,respectively.The expression levels of Rac1 and Cdc42 in CHG5 cells after co-culture were detected using Western blotting. Results: The migration ability of CHG5 co-cultured with GVEC was significantly higher than that in CHG5 co-cultured with VEC and CHG5 culture alone( P <0.05).The results of cytoskeleton immunofluorescence staining showed that the morphology of tubulin in CHG5 cells changed,and the tubulin was thick,circular and radial arrangement after the CHG5 cells were co-cultured with GVEC.The results of Western blotting showed that the expression levels of Rac1 and Cdc42 in CHG5 cells significantly increased after the CHG5 cells were co-cultured with GVEC( P <0.05),while no obvious change of the expression levels of Rac1 and Cdc42 in CHG5 cells was found after the CHG5 cells were co-cultured with VEC ( P > 0.05). Conclusions: The role of tumor cell vascularity is crucial in the migration of glioma cells.The GVEC can promote the migration of tumor cells by changing the cellular tubulin structure,and up-regulating the expression levels of Rac1 and Cdc42 proteins.
Journal of Bengbu Medical College
tumor vascular endothelial cell