胶质瘤细胞对血管内皮细胞迁移的影响  

Influence of clioma cells C6 on migration of endothelial cells

作　　者：林志雄[1] 黄强[2] 杨丽娟[3] 林建华[4] 

机构地区：[1]福建医科大学附属第一医院神经外科,福州350005 [2]苏州大学附属第二医院神经外科 [3]福建医科大学附属第一医院药理学教研室,福州350005 [4]福建医科大学附属第一医院肿瘤侵袭微生态系统研究室,福州350005

出　　处：《中华实验外科杂志》2006年第4期598-600,共3页Chinese Journal of Experimental Surgery

基　　金：福建省科技厅资助项目(01Z034)

摘　　要：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 influence of glioma cells C6 on migration of endothelial cells and its relation with fibronectin （FN） and vascular endothelial growth factor （VEGF）. Methods HUVECs/C6 cells were co-cultured in vitro and the conditioned media were harvested and centrifuged to remove cellular debris, supernatants were placed at 4 ℃ （prepared as chemoattractants for HUVECs and activator for C6 cells）, then the gene transcription and expression of FN in C6 ceils interacted with HUVECs were detected by reverse transcription polymerase chain reaction （RT-PCR） and immunocytochemistry respectively. A rapid in vitro quantitative assay for the migration potential of HUVECs induced by C6 cells was carried out. Selected chemoattractants were treated with neutralizing antibody to FN and to VEGF at 1 mg/L final concentration respectively. Results The down-regulation of transcription and expression of FN was found in C6 cells after co-cultured with HUVECs. The migration of HUVECs in group with 0.1% BSA and RPMI 1640 and C6 cells as chemoattractants was found after 24 h incubation, which could be blocked by VEGF antibody （P 〈 0.05）, but not by FN antibody. No migration was found in the group with both C6 cells and supernatants from co-culture, and the group with only supernatants from co-culture after 24 h incubation. The levels of VEGF in the group with only supernatants from co-cultur as chemoattractants were 0, those in the group with 0.1% BSA and RPMI 1640 and C6 cells were 261. 333 ± 50. 013 ng/L, and those in the group with both C6 cells and supernatants from cocultur were 32. 667 ± 15.011 ng/L, respectively. Conclusion The migration of endothelial cells might be promoted by interactions of glioma cells and endothelial cells, which was contributed to the VEGF.

关 键 词：胶质瘤 纤维连接蛋白 血管内皮生长因子 

分 类 号：R739.4[医药卫生—肿瘤]