利用siRNA抑制HER-2表达的研究
The Inhibition of HER-2 Expression by siRNA
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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。Abstract: 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 transplante...
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