VEGF单抗对低氧环境下C6胶质瘤细胞侵袭性的影响

目的 探讨血管内皮细胞生长因子（VEGF）单克隆抗体对低氧环境下C6胶质瘤细胞迁移和侵袭能力的影响。方法 采用含有10%胎牛血清的DMEM培养基（对照组）培养C6胶质瘤细胞，加入终浓度为100 μmol/L氯化钴模拟缺氧环境（缺氧组），然后加入终浓度为0.1（VEGF-1组）、1（VEGF-2组）、10 μg/ml（VEGF-3组）VEGF抗体处理。MTT法测定细胞活性，Transwell技术检测细胞侵袭和迁移能力，免疫印迹法检测粘着斑激酶（FAK）及富含脯氨酸的酪氨酸激酶2（Pyk2）蛋白及其磷酸化水平。结果 对照组、缺氧组、VEGF-1组和VEGF-2组C6胶质瘤细胞增殖活性无显著差异（P>0.05），但VEGF-3组细胞活性显著降低（P<0.05）。缺氧组和VEGF-1组C6胶质瘤细胞侵袭能力无显著差异（P>0.05），但VEGF-2组C6胶质瘤细胞侵袭能力显著增强（P<0.05）。缺氧组、VEGF-1和VEGF-2组C6胶质瘤细胞FAK蛋白表达及其磷酸化水平无显著差异（P>0.05）。缺氧组、VEGF-1和VEGF-2组C6胶质瘤细胞Pyk2蛋白表达亦无显著差异（P>0.05），但VEGF-2组Pyk2磷酸化水平显著降低（P<0.05）。结论 缺氧条件下，VEGF单抗显著增强C6胶质瘤细胞的侵袭能力，可能与Pyk2磷酸化水平增高有关。

Effect of nonoclonal antibody of VEGF on C6 glioma cells invasiveness under hypoxia and the role of FAK/Pyk2

Objective To investigate the effect of the monoclonal antibody of vascular endothelial growth factor (VEGF)on glioma cell invasiveness under hypoxia and its mechanism. Method C6 glioma cells were treated by VEGF antibodies in vitro under the hypoxia mimicked by CoCl2. C6 glioma cells invasiveness and migration ability were determined respectively by MTT assay, wound healing, and transwell assay. The total and phosphorylated protein levels of focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (Pyk2) were detected by Western blotting. Results The migration ability and invasiveness of C6 glioma cells exposed to VEGF monoantibodies increased in a concentration-dependent manner under the hypoxia. Treatment with VEGF antibodies significantly enhanced the level of Pyk2 phosphorylated at Tyr402, but had no effect on the level of FAK phosphorylated at Tyr397 in vitro. Conclusion The present Results suggest that C6 glioma cell migration ability and invaseness may be promoted anti-VEGF treatment under the hypoxia via the increase in the level of phosphorylated Pyk2 at Pyk402.