利用siRNA抑制食管癌VEGF-C的表达  

Inhibition of VEGF-C Expression in Esophageal Carcinoma by siRNA

作　　者：张红新[1] 郑湘予[1] 陈奎生[1] 张岚[1] 高冬玲[1] 张云汉[1] 

机构地区：[1]郑州大学第一附属医院病理科河南省肿瘤病理重点实验室,郑州450052

出　　处：《肿瘤防治研究》2010年第2期132-135,140,共5页Cancer Research on Prevention and Treatment

基　　金："十五""211工程"重点学科建设基金资助项目(教重办2002第2号);河南省高校杰出科研人才创新工程基金资助项目(2006KYCX016)

摘　　要：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-CmRNA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-CmRNA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-CmRNA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-CmRNA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。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 cells were transplanted into nude mice later. Expression of VEGF-C protein was detected with IHC method in transplanted tumor. Results SiRNA-VEGF-C can decrease the expression of VEGF-C mRNA and protein in EC9706 cells effectively. There were lots of positive granules and positive bands in untransfected EC9706 cells and in the transfection group cells with transfected not-matching any known human coding siRNA. There were few positive granules and week positive bands in transfection group cells with transfected by VEGF-C-targeted siRNA. There were significant differences between transfected and untransfected groups （P〈0.01）. All the three siRNAs targeting VEGF-C could be the tumorgenesis of EC9706 cells in nude mice. Correspondingly, expressions of VEGF-C protein in xenografts tumors were also reduced. There were significant differences between them （P〈0.01）. Conclusion siRNAs targeting VEGF-C can inhibit VEGF-C expression effectively in vivo and in vitro. These maybe contribute to the gene therapy of cancer treatment in human esophageal carcinoma.

关 键 词：VEGF-C 小干扰RNA(siRNA) RT-PCR裸鼠 食管癌 

分 类 号：R735.1[医药卫生—肿瘤]