利用siRNA抑制HER-2表达的研究  

The Inhibition of HER-2 Expression by siRNA

作　　者：吴峰[1] 王清明[1] 范国才[1] 陈吉中[1] 陈惠鹏[2] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]军事医学科学院生物工程研究所

出　　处：《肿瘤防治研究》2006年第4期215-218,298,共5页Cancer Research on Prevention and Treatment

摘　　要：目的利用siRNA抑制人乳腺癌MCF-7细胞中HER-2基因的表达。方法针对HER-2mR-NA设计了5条siRNA,并构建相应的表达质粒,将质粒转染MCF-7乳腺癌细胞,筛选稳定表达株,利用RT-PCR分析siRNA对细胞中HER-2mRNA的降解作用;利用流式细胞术分析细胞表面HER-2蛋白的表达;利用裸鼠进行体内成瘤性实验,用免疫组化法检测瘤组织中HER-2的表达。结果siRNA3P能明显降低MCF-7细胞中HER-2mRNA的丰度;流式细胞分析表明,其中siRNA2P和3P稳定转染的MCF-7细胞表面HER-2的表达明显降低;五种表达不同siRNA的MCF-7细胞在裸鼠体内的成瘤性均有所降低,并且瘤组织中HER-2蛋白的表达也明显减少。结论靶向HER-2mRNA的siRNA能在体内外有效抑制HER-2的表达,可用于乳腺癌的基因治疗。Objective To inhibit the expression of HER-2 gene in MCF-7 human breast carcinoma cells by siRNA. Methods Five siRNAs targeting HER-2 were designed, and their corresponding expression vectors were constructed and transformed into MCF-7 cells. After selection, the stable transfected cells were obtained. The HER-2 mRNA abundance in the stable transfected cells was analyzed by RT-PCR. The HER-2 protein expression in these cells was analyzed by flow cytometry （FCM）. The stable transfected cells were transplanted in nude mice, and the HER-2 protein expression in transplant tumor was detected with immunohistochemical method. Results siRNA 3P could effectively decrease the HER-2 mRNA expression in MCF-7 cells. The FCM data showed that the HER-2 protein expression in siRNA 2P and 3P transfected MCF-7 cells decreased. All the five siRNAs targeting HER-2 could decrease the tumorigenesis of MCF-7 cells in nude mice. Correspondingly, the HER-2 protein expression in transplant tumors was also reduced. Conclusion The siRNAs targeting HER-2 mRNA could effectively inhibit HER-2 expression in vivo and in vitro, and will be useful for gene therapy of breast cancer.

关 键 词：HER-2 小干涉RNA(siRNA) 乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤] Q786[医药卫生—临床医学]