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Abstract：
BRAF gene mutation is found in more than 60% of malignant melanomas, which are difficult to treat. In this study, a new tumor-targeting liposome was developed to deliver anti-BRAF siRNA (siBraf) for the treatment of melanomas. Nucleolin is overexpressed on the surface of cancer cells. AS1411, an aptamer showing specific binding to nucleolin, was conjugated to PEGylated cationic liposome as the targeting probe ASLP (AS1411-PEG-liposome). The ASLP/siRNA complex was formed through electrostatic interaction between ASLP and siRNA. The binding of AS1411 to the surface of PEGylated liposomes was confirmed by gel electrophoresis and capillary electrophoresis. Real-time PCR and Western blot analysis showed that ASLP/siBraf exhibited strong silencing activity of BRAF gene. The much higher accumulation of the siRNA in tumor cells comparing with normal cells indicated that ASLP displayed excellent tumor-targeting capability. Notably, ASLP/siBraf showed significant silencing activity in A375 tumor xenograft mice and inhibited the melanoma growth. These results suggested that the new nucleolin-targeted siRNA delivery system by AS1411 may have the potential for the treatment of melanoma.
摘要：
BRAF基因突变在60%以上的恶性黑色素瘤中发现,很难治疗。在这项研究中,开发了一种新的肿瘤靶向脂质体,用于递送抗BRAFsiRNA(siBraf)治疗黑色素瘤.核仁素在癌细胞表面过度表达。AS1411，一种显示与核仁素特异性结合的适体，与作为靶向探针ASLP(AS1411-PEG-脂质体)的PEG化阳离子脂质体缀合。ASLP/siRNA复合物是通过ASLP和siRNA之间的静电相互作用形成的。通过凝胶电泳和毛细管电泳确认AS1411与聚乙二醇化脂质体表面的结合。实时PCR和Westernblot分析表明，ASLP/siBraf对BRAF基因具有较强的沉默活性。与正常细胞相比,siRNA在肿瘤细胞中的积累高得多,表明ASLP表现出优异的肿瘤靶向能力。值得注意的是,ASLP/siBraf在A375肿瘤异种移植小鼠中显示出显著的沉默活性并抑制黑素瘤生长。这些结果表明，AS1411新的核仁素靶向siRNA递送系统可能具有治疗黑色素瘤的潜力。
