Knock-down of Efp by DNA-modified small interfering RNA inhibits breast cancer cell proliferation and in vivo tumor growth
Knock-down of Efp by DNA-modified small interfering RNA inhibits breast cancer cell proliferation and in vivo tumor growth
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DNA 修饰的小干扰 RNA 敲低 Efp 可抑制乳腺癌细胞增殖和体内肿瘤生长
DOI：
10.1038/cgt.2010.19
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发表时间：
2010
期刊：
影响因子：
6.4
通讯作者：
Inoue S
中科院分区：
文献类型：
--
作者：
Ueyama K;Ikeda K;Sato W;Nakasato N;Horie-Inoue K;Takeda S;Inoue S
雌激素应答基因Efp通过刺激细胞周期负调节因子14-3-3σ的降解来促进乳腺癌细胞的生长，因此被认为是乳腺癌治疗的一个合适的分子靶点。利用小干扰RNA （siRNA）及其衍生物来沉默癌症相关基因正在被研究，目的是确定这些分子的临床应用。近年来，dna修饰的siRNA（嵌合siRNA）在哺乳动物细胞中引起的脱靶效应或免疫反应较少，具有良好的临床应用潜力。在本研究中，我们鉴定出了在MCF-7乳腺癌细胞中沉默Efp表达的最特异性和最有效的siRNA （siEfp-1）。为此，我们使用了一种主要消除脱靶效应的算法。siEfp-1显著抑制胸腺小鼠MCF-7细胞的体外增殖和细胞周期进程，以及MCF-7肿瘤的体内生长。dna修饰的siEfp-1（嵌合siEfp）显著抑制Efp的表达、培养细胞的增殖和mcf -7源性肿瘤在胸腺小鼠体内的生长。此外，siEfp-1和嵌合siEfp沉默Efp表达后，14-3-3σ蛋白的表达增加。这些结果提示siEfp-1和嵌合siEfp可能在乳腺癌治疗中有用。特别是嵌合siEfp，具有高特异性和很少的副作用，因此有望作为一种新的基于核酸的治疗剂。
The estrogen-responsive gene Efp promotes the growth of breast cancer cells by stimulating the degradation of a negative cell-cycle regulator, 14-3-3σ, and is hence considered a suitable molecular target for breast cancer therapy. The use of small interfering RNA (siRNA) and its derivatives to silence cancer-related genes is being investigated with the aim of identifying clinical applications for these molecules. Recently, it has been shown that DNA-modified siRNA (chimeric siRNA) has good potential in clinical applications, because it induces fewer off-target effects or immune responses in mammalian cells. In the present study, we identified the most specific and effective siRNA (siEfp-1) for silencing Efp expression in MCF-7 breast cancer cells. For this purpose, we used an algorithm that primarily eliminates off-target effects. siEfp-1 considerably suppressed the in vitro proliferation and cell-cycle progression of MCF-7 cells, as well as the in vivo growth of MCF-7 tumors, in athymic mice. DNA-modified siEfp-1 (chimeric siEfp) significantly inhibited the expression of Efp, proliferation of cultured cells and the in vivo growth of MCF-7-derived tumors in athymic mice. In addition, the silencing of Efp expression by siEfp-1 and chimeric siEfp increased the expression of the 14-3-3σ protein. These results suggest that siEfp-1 and chimeric siEfp could be useful in breast cancer therapy. Chimeric siEfp, in particular, has a high specificity and induces few side effects and is therefore expected to be used as a novel nucleic acid-based therapeutic agent.