Silencing of the HER2/neu gene by siRNA inhibits proliferation and induces apoptosis in HER2/neu-overexpressing breast cancer cells
Silencing of the HER2/neu gene by siRNA inhibits proliferation and induces apoptosis in HER2/neu-overexpressing breast cancer cells
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DOI：
10.1593/neo.04313
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发表时间：
2004-11-01
期刊：
影响因子：
4.8
通讯作者：
Strebhardt, K
中科院分区：
文献类型：
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作者：
Faltus, T;Yuan, JP;Strebhardt, K
在真核生物中，双链 (ds) RNA 会诱导基因表达的序列特异性抑制，称为 RNA 干扰 (RNAi)。我们利用 RNAi 来定义 HER2/neu 在人类乳腺癌细胞肿瘤增殖中的作用。我们用针对人 HER2/neu 的短干扰 RNA (siRNA) 转染 SK-BR-3、BT-474、MCF-7 和 MDA-MB-468 乳腺癌细胞，并通过 Northern 和 Western blot 分析对 HER2/neu 表达的特异性抑制。用 HER2/neu 特异性 siRNA 转染导致 HER2/neu mRNA 和蛋白质水平出现序列特异性下降。此外，转染 HER2/neu siRNA 导致乳腺癌细胞系 SK-BR-3 和 BT-474 的细胞周期停滞在 G(0)/G(1)，这与强大的 RNA 沉默效应一致。 siRNA 处理在过度表达 HER2/neu 的细胞中产生抗增殖和凋亡反应，但对几乎不表达 HER2/neu 蛋白的细胞（如 MDA-MB-468 细胞）没有影响。这些数据表明 HER2/neu 功能对于 HER2/neu 过表达乳腺癌细胞的增殖至关重要。我们的观察表明，针对人 HER2/neu 的 siRNA 可能是作为抗增殖剂的有价值的工具，在非常低的剂量下就显示出针对肿瘤细胞的活性。
In eukaryotes, double-stranded (ds) RNA induces sequence-specific inhibition of gene expression referred to as RNA interference (RNAi). We exploited RNAi to define the role of HER2/neu in the neoplastic proliferation of human breast cancer cells. We transfected SK-BR-3, BT-474, MCF-7, and MDA-MB-468 breast cancer cells with short interfering RNA (siRNA) targeted against human HER2/neu and analyzed the specific inhibition of HER2/neu expression by Northern and Western blots. Transfection with HER2/neu-specific siRNA resulted in a sequence-specific decrease in HER2/neu mRNA and protein levels. Moreover, transfection with HER2/neu siRNA caused cell cycle arrest at G(0)/G(1) in the breast cancer cell lines SK-BR-3 and BT-474, consistent with a powerful RNA silencing effect. siRNA treatment resulted in an anti proliferative and apoptotic response in cells overexpressing HER2/neu, but had no influence in cells with almost no expression of HER2/neu proteins like MDA-MB-468 cells. These data indicate that HER2/neu function is essential for the proliferation of HER2/neu-overexpressing breast cancer cells. Our observations suggest that siRNA targeted against human HER2/neu may be valuable tools as antiproliferative agents that display activity against neoplastic cells at very low doses.