Implication of the Akt2/survivin pathway as a critical target in paclitaxel treatment in human ovarian cancer cells
Implication of the Akt2/survivin pathway as a critical target in paclitaxel treatment in human ovarian cancer cells
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Akt2/survivin 通路作为紫杉醇治疗人卵巢癌细胞关键靶点的意义
DOI：
10.1016/j.canlet.2008.08.027
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发表时间：
2009-01-18
期刊：
影响因子：
9.7
通讯作者：
Ma, Ding
中科院分区：
文献类型：
--
作者：
Weng, Danhui;Song, Xiaohong;Ma, Ding
目的：尽管紫杉醇（PTX）诱导的卵巢癌耐药涉及多种机制，但最近的证据表明，Akt 2在保护细胞免受紫杉醇诱导的凋亡中具有重要作用。在本研究中，我们研究了Akt 2/survivin通路在紫杉醇诱导的耐药中的作用，通过一种改进的方法来产生一个有效的shRNAvector.Methods：我们应用RNAi介导的沉默技术来研究Akt 2/survivin通路在M诱导的卵巢癌细胞（A2780和SKOV 3）耐药中的机制。采用半定量实时荧光定量RT-PCR和western blot检测Akt 2和survivin mRNA及相关蛋白的表达水平。MTT法检测Akt 2对卵巢癌细胞增殖的抑制作用，流式细胞仪检测Akt 2对卵巢癌细胞凋亡的诱导作用，Hoechst染色检测Akt 2对卵巢癌细胞凋亡的诱导作用。我们进一步证实抑制survivin表达的抑制可以诱导紫杉醇耐药。我们引入了一个改进的载体，产生shRNA诱导RNA干扰，其中包含三个U6启动子表达不同的shRNA：它严重降低Akt 2基因的表达，并表现出良好的特异性。结论：我们的研究结果将有助于了解紫杉醇诱导卵巢癌耐药的分子机制，并促进新的抗肿瘤策略的发展。(C)2008爱思唯尔爱尔兰有限公司保留所有权利。
Purpose: Although multiple mechanisms have been implicated in paclitaxel (PTX)-induced resistance in ovarian cancer, recent evidence has suggested that Akt2 has an important role in the protection of cells from paclitaxel-induced apoptosis. In the present study, we investigated the role of the Akt2/survivin pathway in paclitaxel-induced resistance by a modified method to generate an effective shRNA vector.Methods: We applied RNAi-mediated silencing techniques to investigate the mechanism of the Akt2/survivin pathway on M-induced resistance in ovarian cancer cells (A2780 and SKOV3). The expression of Akt2 and survivin mRNA and related protein levels were evaluated with semiquantitative real-time RT-PCR and western blot analysis, respectively. Inhibition of cell proliferation was determined by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay, and the induction of apoptosis was examined through flow cytometry (FACS) and Hoechst staining.Results: Akt2 down-regulation sensitized ovarian cancer cells to paclitaxel-induced apoptosis, and inhibited survivin expression. We further demonstrated that suppressing the inhibition of survivin expression can induce the drug-resistance to paclitaxel. We introduced a modified vector to generate shRNA to induce RNA interference, which contained three U6 promoters to express different shRNAs: it severely reduced Akt2 gene expression and showed good specificity.Conclusion: Our findings will aid in understanding the molecular mechanism of paclitaxel-induced resistance in ovarian cancer and facilitate the development of novel anti-neoplastic strategies. (C) 2008 Elsevier Ireland Ltd. All rights reserved.