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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 inhibitory effect of specific small interfering RNA (siRNA) of vascular endothelial growth factor (VEGF) on VEGF expression in poorly differentiated human nasopharyngeal carcinoma CNE-2Z cells and to provide a new approach for the treatment of nasopharyngeal carcinoma. Methods Human nasopharyngeal carcinoma cell line CNE-2Z was cultured in vitro and divided into normal oxygen group (20%) and hypoxic (1%) group. VEGF siRNA was transfected into two groups of cells by lipofectamine2000 (LF2000), and RT-PCR and ELISA were used to detect the difference of VEGF mRNA and protein expression between nasopharyngeal carcinoma cells under normal oxygen and hypoxia. Results Compared with untransfected group and empty vector transfected group, VEGFsiRNA significantly inhibited the expression of VEGFmRNA (P <0.01) under normoxia and hypoxia. The inhibitory efficiency of VEGFsiRNA in normoxia was lower than that in hypoxia high. Under normal oxygen conditions, the expression of VEGF165 in CNE-2Z cells transfected with VEGFsiRNA group was 423.92, 419.68 and 328.86 pg / mL, respectively. The expression of VEGF165 in hypoxia The content of VEGF was 813.62, 799.89 and 500.96 pg / mL, respectively. The expression of VEGF in transfected VEGFsiRNA group was down-regulated. Conclusion VEGFsiRNA can effectively and specifically inhibit the expression of VEGFmRNA and protein in human nasopharyngeal carcinoma CNE-2Z cells, which is expected to be an effective method for the treatment of nasopharyngeal carcinoma.