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目的探讨胶质瘤细胞对血管内皮细胞迁移行为的影响。方法利用体外快速侵袭力测定法检测C6细胞对人脐静脉内皮细胞(HUVECs)迁移的影响;ELISA方法分析迁移诱导剂中血管内皮细胞生长因子(VEGF)的浓度;逆转录-聚合酶链反应(RT-PCR)和免疫细胞化学方法检测 C6细胞上纤维连接蛋白(FN)的基因转录及蛋白的表达情况。结果以C6细胞和HUVECs的共培养上清液为HUVECs迁移诱导剂组及以C6细胞+共培养上清液为诱导剂组孵育24 h后未见 HUVECs迁移,而以0．1％BSA+RPMI 1640+C6细胞为诱导剂组孵育24 h后可见HUVECs迁移,且这种迁移可被VEGF抗体所封闭(P<0．05),但不被FN抗体所封闭;单纯共培养上清液组、 0．1％BSA+RPMI 1640+C6细胞组及C6细胞+共培养上清液组的VEGF浓度分别为0 ng／L, (261．333±50．013)ng／L,(32．667±15．011)ng／L;C6细胞经和HUVECs共培养后出现FN的 mRNA转录和蛋白表达的下调。结论 C6细胞通过VEGF的作用促进血管内皮细胞向肿瘤细胞迁移。
Objective To investigate the effect of glioma cells on migration of vascular endothelial cells. Methods The effect of C6 cells on the migration of human umbilical vein endothelial cells (HUVECs) was assayed by rapid invasiveness assay in vitro. The levels of vascular endothelial growth factor (VEGF) in the migration inducer were analyzed by ELISA. The levels of VEGF, RT-PCR and immunocytochemistry were used to detect the gene transcription and protein expression of fibronectin (FN) on C6 cells. Results HUVECs migration was not observed when the co-culture supernatant of C6 cells and HUVECs were HUVECs migration inducer and C6 cells plus co-culture supernatants as inducer for 24 h. However, HUVECs migrated with 0.1% BSA + RPMI The migration of HUVECs was observed after incubation with 1640 + C6 cells for 24 h as inducer, and the migration was blocked by VEGF antibody (P <0.05), but not blocked by FN antibody. In pure co-culture supernatant group, The concentrations of VEGF in 0.1% BSA + RPMI 1640 + C6 cells and C6 cells + co-culture supernatants were 0 ng / L, (261.333 ± 50.013) ng / L and 15.011) ng / L. Down-regulation of mRNA and protein expression of FN was observed in C6 cells co-cultured with HUVECs. Conclusion C6 cells promote the migration of vascular endothelial cells to tumor cells through the action of VEGF.