RNA干扰抑制结肠癌血管内皮生长因子的体内实验研究

Inhibit Vegf Expression of Colon Cancer by Rna Interference in Vivo

目的:应用RNA干扰技术抑制血管内皮生长因子(VEGF)表达,并通过体内实验,观察在体内复杂环境下能否有效抑制VEGF表达,以及结肠癌细胞凋亡的情况,为RNA干扰技术的临床应用奠定基础。方法:体内实验中,将RNA干扰质粒以水动力法导入结肠癌裸鼠模型,通过RT-PCR观察VEGF mRNA表达受抑的程度;Western杂交检测VEGF蛋白的表达情况;通过测量比较瘤体大小和Tunel法,观察VEGF受抑制后,肿瘤形态和细胞凋亡的变化。结果:体内实验中,通过RT-PCR和Western杂交检测,发现这两种抗VEGF表达载体能够有效抑抑制裸鼠结肠癌模型瘤体VEGF的表达;VEGF受抑制后,瘤体增长受到抑制,Tunel法显示,RNA干扰可致肿瘤细胞凋亡增加。结论:Tunel法分析显示,当VEGF受抑制后,肿瘤细胞凋亡发生率明显增加,针对VEGF基因的shRNA表达载体能够明显抑制裸鼠VEGF基因的表达。

To inhibit VEGF expression by RNA interference in vivo,and observe the apoptosis of Colon cancer when VEGF was knocked down,so that RNA interference could be used in clinical therapy.Methods：Injecting the vectors into nude mouse modle of Colon cancer through tail vein by Hydrodynamics methods,the expression of VEGF was investigated by RT-PCR and Western blot.When the level of VEGF was knocked down,the size of tumor body was measured,and the apoptosis of tumor cells was investigated by Tunel.Results：the similar results were also got in vivo,showed the expression of VEGF could be efficiently inhibited in nude mouse model analyzed by RT-PCR.When the level of VEGF was knocked down,the size of tumor body was reduced and the apoptosis of tumor cells were increased showed by Tunel. Conclusion：The expression of VEGF could be inhibited by RNA interference in vivo.The inhibition of VEGF could induce the apoptosis of tumor cells in vivo.