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目的探讨诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)基因对舌鳞癌细胞株 Tca8113细胞中血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达调控情况。方法以 RNA 干扰技术将含有 iNOS 基因的特异性(short hairpin RNA,shRNA)片断的质粒表达载体转染入 Tca8113细胞中,RT-PCR 和 Western blot 法分别检测 iNOS、VEGF 的 mRNA、蛋白表达改变情况。结果 Tca8113细胞的 iNOS 对 VEGF 基因的 PCR 产物,在转染后24、48 h 实验组和对照组之间比较,差异均有统计学意义(P<0.05);iNOS 基因的蛋白产物在转染后36、48 h 差异均有统计学意义(P<0.05)。VEGF 基因的蛋白产物在转染后48 h 差异有统计学意义(P<0.05)。结论以 RNA 干扰技术沉默 Tca8113细胞中 iNOS 基因,可以降低细胞中 VEGF 的表达水平,iNOS 基因可能成为肿瘤防治的新靶点。
Objective To investigate the regulatory effect of inducible nitric oxide synthase (iNOS) gene on the expression of vascular endothelial growth factor (VEGF) in tongue squamous carcinoma cell line Tca8113. Methods Plasmid expression vector containing short hairpin RNA (shRNA) fragment of iNOS gene was transfected into Tca8113 cells by RNA interference technique. The changes of mRNA and protein expression of iNOS and VEGF were detected by RT-PCR and Western blot respectively . Results The iNOS gene of Tca8113 cells showed a significant difference (P <0.05) in the PCR products of VEGF gene between the experimental group and the control group 24 and 48 h after transfection (P <0.05). The protein products of iNOS gene after transfection 36,48 h difference was statistically significant (P <0.05). The protein expression of VEGF gene at 48 h after transfection was significantly different (P <0.05). Conclusion The silencing of iNOS gene in Tca8113 cells by RNA interference can reduce the expression of VEGF in the cells. The iNOS gene may be a new target for tumor prevention and treatment.