利用siRNA抑制食管癌VEGF-C的表达

目的 利用siRNA抑制人食管癌EC9706细胞中VEGF-C基因的表达。方法 针对VEGF-C mRNA设计了3条siRNA，并构建相应的表达质粒，将质粒转染食管癌EC9706细胞，筛选稳定表达株，利用RT-PCR和原位杂交技术分析siRNA对细胞中VEGF-C mRNA的降解作用；免疫组织化学法分析细胞中VEGF-C蛋白的表达；利用裸鼠进行体内成瘤性实验，免疫组织化学法检测瘤组织中VEGF-C的表达。结果 siRNA能明显降低食管癌EC9706细胞中VEGF-C mRNA和蛋白的表达，稳定转染的3个siRNA组细胞仅有少量VEGF-C阳性表达颗粒和无阳性条带，与无关序列组和正常EC9706细胞组相比，差异均有统计学意义（P<0.01）；裸鼠体内的成瘤实验，3个siRNA转染组瘤组织中VEGF-C蛋白的表达也明显减少，与正常EC9706组无关序列组相比，差异均有统计学意义（P<0.01）。结论 靶向VEGF-C mRNA的siRNA能在体内外有效抑制VEGF-C的表达，可用于食管癌的基因治疗。

Inhibition of VEGF-C Expression in Esophageal Carcinoma by siRNA

Objective To inhibit the expression of VEGF-C gene in human esophageal carcinoma cells EC9706 by siRNA. Methods Three siRNAs targeting VEGF-C were designed. Corresponding expression vectors were constructed and transformed into EC9706 cells. After selection,the stable transfected cells were obtained. VEGF-C mRNA in the stable transfected cells was analyzed by RT-PCR and in situ hybridization. Expression of VEGF-C protein in these cells was analyzed by immunohistochemistry (IHC) method. Stable transfected ce...