VHL基因对U251胶质瘤细胞侵袭和迁移的影响

  目的   研究VHL基因对U251胶质瘤细胞侵袭和迁移能力的影响   方法   使用VHL表达质粒转染胶质瘤细胞，采用RT-PCR检测VHL mRNA表达，Western blot检测VHL、MMP-2、MMP-9蛋白表达，应用Transwell体外侵袭实验、划痕实验检测上调VHL基因后对U251胶质瘤细胞侵袭迁移能力的影响。使用VHL表达质粒处理后的U251细胞建立裸鼠颅内模型，利用免疫组织化学染色法对颅内模型组织切片中VHL、MMP-2、MMP-9蛋白表达进行检测。   结果   VHL表达质粒转染胶质瘤细胞后VHL mRNA、VHL蛋白表达增高，MMP-2、MMP-9蛋白表达下降；并抑制U251胶质瘤细胞侵袭迁移能力。对染色后的组织切片进行分析，VHL表达质粒处理组中VHL的表达较对照组明显升高，MMP-2、MMP-9蛋白较对照组表达减少。   结论   VHL基因能够有效抑制U251胶质瘤细胞侵袭迁移能力，VHL基因可成为治疗胶质瘤的一个有效靶点。 

Effect of VHL on the invasion and migration of glioma U251 cells

  Objective   To determine the effects of Von Hippel-Lindau (VHL) on the invasion and migration of glioma U251 cells.   Methods   U251 GBM cells were transfected using VHL expression plasmid. Real-time polymerase chain reaction was conducted to detect VHL mRNA expression after transfection. Western blot assay was used to measure protein (VHL, MMP-2, and MMP-9) expression. Tumor invasion and migration were examined by the Transwell and wound-healing experimental methods after VHL up-regulation. The intracranial model of nude mouse was developed using U251 cells transfected by VHL expression plasmid, and immunohistochemical staining was used to measure protein (VHL, MMP-2, and MMP-9) expression in the tissue sections.   Results   In the U251 cells transfected by VHL expression plasmid, the expression of VHL mRNA and VHL proteins increased, and the expression of MMP-2 and MMP-9 protein decreased. Meanwhile, the invasion and migration of glioma U251 cells were also inhibited. Immunohistochemical staining results showed that the expression of MMP-2 and MMP-9 proteins decreased, and the VHL protein expression increased after transfection.   Conclusion   VHL can inhibit the invasion and migration of glioma U251 cells. Thus, VHL gene can be used as a target for the gene therapy of gliomas.