摘要
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VEGFsiRNA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VEGFmRNA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VEGFsiRNA组VEGF的表达下调。结论VEGFsiRNA能有效地、特异地抑制人鼻咽癌CNE-2Z细胞VEGFmRNA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。
[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% 02） and the hypoxia （1% 02） culture group. VEGFsiRNA were transfected into CNE-2Z ceils 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; Downregnlation 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）.
出处
《中国医学工程》
2007年第3期231-234,共4页
China Medical Engineering
基金
国家自然科学基金资助项目(30400118)
关键词
RNA干扰
血管内皮生长因子
鼻咽癌
基因治疗
RNA interference
Vascular endothelial growth factor
Nasopharyngeal Cancer cells
Gene therapy
