小干扰RNA对人胰腺癌细胞移植瘤血管生成的影响

The influence of VEGF siRNA on the angiogenesis of human pancreatic cancer( PANC-1) cells transplantation tumor

目的研究小干扰RNA(small interfering RNA,siRNA)抑制血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达对裸鼠皮下人胰腺癌PANC-1细胞移植瘤血管生成的影响。方法构建人胰腺癌PANC-1细胞40只裸鼠皮下移植瘤模型,随机分为4组。两组针对VEGF的siRNA真核表达载体(PU-VEGF-siRNA1,PU-VEGF-siRNA2)通过脂质体法行瘤内和瘤周注射;注射空质粒组作为实验对照组;未注射组作为空白对照组。注射1次/3天,共10次,末次注射3天后,剥离瘤体,免疫组化染色法观察VEGF阳性染色及微血管参数,RT-PCR检测瘤组织VEGF-mRNA表达。结果 PU-VEGFsiRNA2组移植瘤VEGF-mRNA及转染PU-VEGF-siRNA后肿瘤组织微血管密度(MVD)表达均较空质粒组及空白对照组低(P<0.05)。而PU-VEGF-siRNA1组未发现差别(P>0.05)。结论 siRNA能在体内抑制胰腺癌VEGF表达,抑制肿瘤血管生成。

Objective To investigate the influence of VEGFsiRNA on the angiogenesis of human pancreatic cancer（ PANC-1） cells transplantation tumor. Methods Human pancreatic cancer tumors were established on the backs of 40 nude mice by subcutaneous injection of cultured cells（ PANC-1）. The animals were divided into4 groups randomly. Two siRNA targeting VEGF constructed in eukaryotic expression vector（ PU-VEGF-siRNA1,PU-VEGF-siRNA2） were given intratumoral and peritumoral injection with lipofectamine respectively. Tumors injected vector without siRNA and non-injected tumors were used as experimental and negative control groups.Animals were injected one time every 3 days for total 10 times. Three days after the last injection,the tumors were excised,and the expression of VEGF in xenograft tumors were detected by reverse transcription polymerse chain reaction（ RT-PCR）. The protein expression of VEGF and microvessel density（ MVD） of tumor tissue were detected by immunohistochemical technique. Results Compared to the experimental and negative control groups,the expression of VEGF-mRNA and MVD were both decreased（ P 0. 05）. But there was no significantly difference in PU-VEGF-siRNA1 group（ P 0. 05）. Conclusions siRNA can reduce the expression of VEGF in vivo,and inhibit the tumor angiogenesis.