摘要
目的探讨siRNA沉默血管内皮生长因子(VEGF)基因对人肾癌裸鼠移植瘤VEGF蛋白的表达水平及生长抑制作用的影响。方法化学合成针对VEGF的siRNA序列,通过脂质体将VEGF-siRNA转染到ACHN细胞中,将转染VEGF-siRNA的ACHN细胞(A组)、转染空质粒的ACHN细胞(B组)及未转染的ACHN细胞(C组)分别接种于裸鼠背部皮下。在SPF环境中饲养裸鼠并观察各组裸鼠移植瘤的生长情况,每隔5d测定移植瘤体积大小(肿瘤的长轴L,短轴W,肿瘤体积V=1/2LW2),绘制肿瘤生长曲线,采用免疫组化及Western blot法测定裸鼠移植瘤组织切片中VEGF蛋白的表达水平。结果 A组小鼠移植瘤成瘤及生长明显缓慢,移植瘤的体积和质量均低于B、C组,差异有统计学意义(P<0.05);移植瘤组织切片免疫组化及Western blot法检测结果提示:与B、C组相比,A组中VEGF蛋白表达量降低,差异有统计学意义(P<0.05);而B、C组间瘤体的体积重量及VEGF蛋白表达差异均无统计学意义(P>0.05)。结论 VEGF在肾癌的发生、发展中起着重要作用,化学合成的VEGF-siRNA可特异性抑制肾癌细胞中VEGF的表达,抑制肿瘤的生长增殖。
Objective To investigate the effect of siRNA vascular endothelial growth factor（VEGF） gene silencing on the VEGF protein expression level and tumor development in a xenograft model of renal cell carcinoma.Methods The VEGF-siRNA sequence was chemosynthezed and selected in vitro in the previous experiments.The ACHN cells were transfected with selected VEGF-siRNA in Group A;transfected with plasmid DNA in Group B,and without transfection in Group C using liposome as vector.The nude mice were raised in SPF environment,and were observed on development of the transplanted tumor.The tumor dimension（length L,width W,and the volume V=1/2 LW2） was recorded every 5 days,while the tumor growth curve was drawn at the same time to show the inhibitory effects of siRNA on the tumor growth.The VEGF protein expression in the tissue section of the transplanted tumor in mice was also tested through immunohistochemistry method and Western blot test was used to analyze the silencing effects of siRNA on VEGF expression.Results The transplanted tumor developed and grew slowly in Group A.The tumor volume and weight of Group A were significantly lower than Group B and C（P〈0.05）.The VEGF protein expression in Group A was significantly inhibited as indicated by the immunohistochemistry test and Western blot test（P〈0.05）,while the Group B and Group C showed no significant difference in neither tumor volume and weight or VEGF protein expression（P〈0.05）.Conclusion The VEGF plays a important role in the development and growth of renal cancer.The VEGF-siRNA showed specific inhibitory effect on the VEGF expression and tumor development in a xenograft model of renal cell carcinoma.
出处
《热带医学杂志》
CAS
2011年第11期1230-1233,F0003,共5页
Journal of Tropical Medicine
基金
安徽省高校省级自然科学研究项目(ZD200907)
关键词
小干扰RNA
血管内皮生长因子
裸鼠
移植瘤
肾癌
siRNA
vascular endothelial growth factor
nude mice
transfected tumor
kidney cancer
