Li Yuhuan, Liu Da, Zhou Yulin, Li Yujing, Xie Jing, Lee Robert J, Cai Yong, Teng Lesheng
1. Institute of Life Sciences, Jilin University, Changchun, Jilin, P. R. China.
1. Institute of Life Sciences, Jilin University, Changchun, Jilin, P. R. China ; 2. Division of Pharmaceutics, College of Pharmacy, The Ohio State University, Columbus, OH, U.S.A.
J Cancer. 2015 Sep 15;6(11):1187-94. doi: 10.7150/jca.12437. eCollection 2015.
Survivin is an anti-apoptotic gene that is overexpressed in most human tumors. RNA interference using short interfering RNA (siRNA) can be used to specifically inhibit survivin expression. Tumor cells were treated with a newly designed survivin siRNA, which was modified with 2'-OMe. Cellular survivin mRNA and protein levels were determined by real-time qRT-PCR and Western blot, respectively. Cell cycle and apoptosis were determined by flow cytometry. Cell proliferation was measured by MTT assay. Our data showed that the novel survivin-targeted siRNA could efficiently knockdown the expression of survivin and inhibit cell proliferation. Survivin mRNA was reduced by 95% after 48h treatment with 20nM siRNA. In addition, the siRNA could markedly arrest the cell cycle at the G2/M checkpoint and induce cellular apoptosis in a dose-dependent manner. The percentage of apoptotic cells reached 50% when treated with 40nM siRNA. In conclusion, we have identified a novel chemically modified siRNA against survivin that is highly efficient and delineated its mechanism of action, thus demonstrating a potential therapeutic role for this molecule in cancer. Further evaluation of this siRNA for therapeutic activity is warranted.
生存素是一种抗凋亡基因，在大多数人类肿瘤中过度表达。使用小干扰RNA（siRNA）的RNA干扰可用于特异性抑制生存素的表达。用新设计的经2'-O-甲基修饰的生存素siRNA处理肿瘤细胞。分别通过实时定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法测定细胞内生存素mRNA和蛋白质水平。通过流式细胞术测定细胞周期和细胞凋亡。通过MTT法测量细胞增殖。我们的数据表明，新型靶向生存素的siRNA能够有效敲低生存素的表达并抑制细胞增殖。用20nM siRNA处理48小时后，生存素mRNA降低了95%。此外，siRNA能够以剂量依赖的方式在G2/M检查点显著阻滞细胞周期并诱导细胞凋亡。用40nM siRNA处理时，凋亡细胞百分比达到50%。总之，我们鉴定出一种针对生存素的新型化学修饰siRNA，其效率很高，并阐明了其作用机制，从而证明了该分子在癌症中的潜在治疗作用。有必要对这种siRNA的治疗活性进行进一步评估。
