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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-silenced VEGF gene on the expression of VEGF protein and the growth inhibitory effect on human renal cell carcinoma in nude mice. Methods siRNA targeting siRNA was synthesized and transfected into ACHN cells by liposome. ACHN cells transfected with VEGF-siRNA (group A), ACHN cells transfected with empty vector (group B) and Untransfected ACHN cells (group C) were inoculated subcutaneously in nude mice respectively. The growth of nude mice xenografts was observed in nude mice in each SPF group. The volume of tumor xenografts (long axis L, short axis W, tumor volume V = 1 / 2LW2) was determined every 5 days, and the tumor growth Curve, immunohistochemistry and Western blot were used to detect the expression of VEGF protein in the xenografts of nude mice. Results The tumorigenicity and growth of the transplanted tumor in group A were obviously slower than those in group B and C (P <0.05). The tumor tissue sections were immunohistochemically and Western blot The results showed that the expression of VEGF protein in group A was significantly lower than that in group B and C (P <0.05), but there was no difference between the volume and VEGF protein expression in group B and C Statistical significance (P> 0.05). Conclusion VEGF plays an important role in the development and progression of renal cell carcinoma. Chemically synthesized VEGF-siRNA can specifically inhibit the expression of VEGF in renal cell carcinoma and inhibit the growth and proliferation of the tumor.