- 作者：Danhui Weng ; Xiaohong Song ; Hui Xing ; Xiaoli Ma ; Xi Xia ; Yanjie Weng ; Jianfeng Zhou ; Gang Xu ; Li Meng ; Tao Zhu ; Shixuan Wang ; Ding Ma
- 关键词：Akt2 ; Survivin ; shRNA ; Paclitaxel ; Chemoresistance ; Apoptosis
- 刊名：Cancer Letters
- 出版年：2009
- 出版时间：18 January 2009
- 年：2009
- 卷：273
- 期：2
- 页码：257-265
- 全文大小：1209 K
PurposeAlthough 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 PTX-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.