血管内皮生长因子干扰RNA对人鼻咽癌CNE一2Z细胞VEGF表达调控的实验研究

目的：研究血管内皮生长因子（VEGF）的特异性小片段干扰RNA（siRNA）对低分化人鼻咽癌CNE-2Z细胞VEGF表达的抑制作用，为鼻咽癌的治疗提供新的途径。方法：体外培养人鼻咽癌细胞CNE-2Z，并分成正常氧状态培养组（20％）和低氧（1％）培养组。采用脂质体（LF2000）将VEGF siRNA转染两组细胞，并采用RT—PCR、ELISA检测鼻咽癌细胞在正常氧状态和低氧状态下VEGF mRNA和蛋白质表达的差异。结果：与未转染组和转染空载体组相比，在正常氧和低氧状态下，VEGFsiRNA均能明显抑制VEGFmRNA的表达（P〈0．01）；正常氧状态下VEGFsiRNA的抑制效率比低氧状态高。正常氧状态下：未转染组、转染空载体组、转染VEGFsiRNA组CNE-2Z细胞培养上清液中VEGF165的含量分别为：423．92、419．68和328．86pg／mL；低氧状态下VEGF165的含量分别为：813．62、799．89和500．96pg／mL，转染VEGF siRNA组VEGF的表达下调。结论：VEGF siRNA能有效地、特异地抑制人鼻咽癌CNE-2Z细胞VEGF mRNA和蛋白质的表达，因而有望成为治疗鼻咽癌的一种有效方法。

A small interfering RNA targeting vascular endothelial growth factor inhibits CNE-2Z nasopharyngeal cancer cell neovascularization in vitro

Objective] To study the inhibiting effect of vascular endothelial growth factor (VEGF) siRNA to the expression of VEGF in CNE-2Z Nasopharyngeal Cancer cells, and to explore the feasibility of VEGFsiRNA in treating nasopharyngeal cancer. Methods] Nasopharyngeal squamous cells of human (CNE-2Z) were cultured and divided into the normoxia culture group (20% O2) and the hypoxia (1% O2) culture group. VEGFsiRNA were transfected into CNE-2Z cells of two groups with liposome (LF2000). VEGF mRNA expressions of normoxia and hypoxia were tested by RT-PCR and ELISA assay. Results] VEGF mRNA was expressed in CNE-2Z cells cultured in normoxia. VEGFmRNA expression increased in hypoxia, and there is significant difference between the two groups (P <0.01). VEGFsiRNA downregulated VEGFmRNA expression significantly compared with that of non-transfection group and the vector transfection group (P <0.01) both in normoxia and hypoxia; Downregulation of VEGFmRNA expression in normoxia was higher than in hypoxia. Conclusion] VEGF-specific siRNA can inhibit VEGF mRNA expression in nasopharyngeal squamous cell of human (CNE-2Z).