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目的:观察小干扰RNA(small interfering RNA,siRNA)抑制血管内皮生长因子(vascular endothelial growthfactor,VEGF)表达对裸鼠皮下人舌癌Tca8113细胞移植瘤血管生成的影响。方法:构建人舌癌Tca8113细胞20只裸鼠皮下移植瘤模型,随机分为4组。将两对针对VEGF的siRNA真核表达载体(Pu-VEGF-siRNA1,Pu-VEGF-siRNA2)脂质体法作瘤体内及瘤周注射;以注射空质粒组作实验对照组;未注射组作空白对照组。注射1次/3d×10次,末次注射3d后,剥离瘤体,RT-PCR检测瘤组织VEGF-mRNA表达,免疫印迹技术检测瘤组织VEGF蛋白表达。免疫组化染色法观察VEGF阳性染色及微血管参数。结果:Pu-VEGF-siRNA2组移植瘤VEGF-mRNA表达及VEGF蛋白表达、总体微血管密度、有腔微血管密度、有腔血管平均血管周长及管腔面积均较空质粒组及空白对照组低(p<0.05)。而Pu-VEGF-siRNA1组未观察到上述差别(p>0.05)。结论:siRNA能在体内抑制舌癌VEGF表达,有效减少肿瘤血管生成。不同的干扰片段体内作用效果存在差异。
OBJECTIVE: To observe the effects of small interfering RNA (siRNA) on the angiogenesis of subcutaneous human tongue cancer Tca8113 cells in nude mice by inhibiting the expression of vascular endothelial growth factor (VEGF). METHODS: Twenty nude mice xenografts of tongue cancer Tca8113 cells were subcutaneously transplanted into nude mice and were randomly divided into 4 groups. Two pairs of VEGF eukaryotic expression vectors (Pu-VEGF-siRNA1, Pu-VEGF-siRNA2) were injected intratumoral and peritumoral in vivo. Blank control group. The tumor was dissected 1 / 3d × 10 times. After the last injection, the tumor was dissected and the expression of VEGF-mRNA was detected by RT-PCR. The expression of VEGF protein was detected by immunoblotting. Immunohistochemical staining was used to observe VEGF positive staining and microvascular parameters. Results: The expression of VEGF-mRNA, VEGF protein, total microvessel density, luminal microvessel density, aortic mean vessel length and lumen area in Pu-VEGF-siRNA2 group were lower than those in blank plasmid group and blank control group p <0.05). The above difference was not observed in Pu-VEGF-siRNA1 group (p> 0.05). Conclusion: siRNA can inhibit VEGF expression in tongue cancer in vivo, and effectively reduce tumor angiogenesis. The effect of different interference fragments in vivo effects vary.