Imaging Dendrimer-Grafted Graphene Oxide Mediated Anti-miR-21 Delivery With an Activatable Luciferase Reporter
Imaging Dendrimer-Grafted Graphene Oxide Mediated Anti-miR-21 Delivery With an Activatable Luciferase Reporter
复制标题
使用可激活荧光素酶报告基因对树枝状聚合物接枝氧化石墨烯介导的抗 miR-21 递送进行成像
DOI：
10.1021/acsami.6b02662
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发表时间：
2016
影响因子：
9.5
通讯作者：
Tian Jie
中科院分区：
文献类型：
--
作者：
Wang Fu;Zhang Beilei;Zhou Lin;Shi Yaru;Li Zhiqiang;Xia Yuqiong;Tian Jie
MicroRNAs(MiRNAs)是一类转录后基因调控因子，参与肿瘤发生等多种生理过程，已成为肿瘤治疗的潜在靶点。然而，由于缺乏有效的递送载体，被称为抗miRs的反义寡核苷酸在体内用于拮抗miRNA功能的应用在很大程度上受到了阻碍。在这里，我们描述了聚酰胺胺(PAMAM)树状大分子和聚乙二醇化的纳米氧化茂(NGO)偶联物(NGO-PEG-Dendrimer)的发展，以有效地将抗miR-21转移到非小细胞肺癌细胞中。为了监测抗miR-21进入细胞和肿瘤的情况，我们还构建了一个可激活的荧光素酶报告基因(Fluc-3xPS)，在该报告的3‘非翻译区(UTR)中包含三个针对miR-21的完全互补的序列。与未修饰的树枝状大分子和脂质体2000(Lipo2000)相比，非政府组织-聚乙二醇树状大分子具有较低的细胞毒性和较高的转染率。体外生物发光成像和Western blotting分析表明，NGO-PEG-Dendrimer有效地将抗miR-21基因导入细胞质，并以剂量依赖的方式上调荧光素酶强度和PTEN靶蛋白的表达。此外，非政府组织-聚乙二醇-树状大分子转染抗miR-21对细胞迁移和侵袭的抑制作用强于裸树突状大分子或Lipo2000。通过非政府组织-聚乙二醇树状大分子静脉注射抗miR-21可以显著增加报告移植瘤区域内的生物发光信号。这些结果表明，非政府组织-聚乙二醇-树枝状大分子可能是一种有效的、潜在的运送RNA寡核苷酸的纳米载体。此外，将非政府组织-聚乙二醇-树枝状大分子与可激活的荧光素酶报告相结合的策略允许对传递过程进行图像引导监测，这可以为基于RNA的癌症治疗提供见解。
MicroRNAs (miRNAs) are a class of post-transcriptional gene regulators involved in various physiological processes including carcinogenesis, and they have emerged as potential targets for tumor theranostics. However, the employment of antisense oligonucleotides, termed anti-miRs, for antagonizing miRNA functions in vivo has largely been impeded by a lack of effective delivery carriers. Here, we describe the development of polyamidoamine (PAMAM) dendrimer and polyethylene glycol (PEG)-functionalized nanographene oxide (NGO) conjugate (NGO-PEG-dendrimer) for the efficient delivery of anti-miR-21 into non-small-cell lung cancer cells. To monitor the delivery of anti-miR-21 into cells and tumors, we also constructed an activatable luciferase reporter (Fluc-3xPS) containing three perfectly complementary sequences against miR-21 in the 3′ untranslated region (UTR) of the reporter. Compared with bare dendrimer and Lipofectamine 2000 (Lipo2000), NGO-PEG-dendrimer showed considerably lower cytotoxicity and higher transfection efficiency. As demonstrated by in vitro bioluminescence imaging and Western blotting assays, NGO-PEG-dendrimer effectively delivered anti-miR-21 into the cytoplasm and resulted in the upregulation of luciferase intensity and PTEN target protein expression in a dose-dependent manner. Moreover, transfection with anti-miR-21 by NGO-PEG-dendrimer led to stronger inhibition of cell migration and invasion than did bare dendrimer or Lipo2000 transfection. The intravenous delivery of anti-miR-21 via NGO-PEG-dendrimer induced a significant increase in the bioluminescence signal within the Fluc-3xPS reporter-transplanted tumor areas. These results suggest that NGO-PEG-dendrimer could be an efficient and a potential nanocarrier for delivering RNA oligonucleotides. In addition, the strategy of combining NGO-PEG-dendrimer with an activatable luciferase reporter allows the image-guided monitoring of the delivery process, which can provide insights into the RNA-based cancer treatments.