Engineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University , Xi'an, Shaanxi 710071, China.
Department of Gynecology and Obstetrics, Tangdu Hospital, The Fourth Military Medical University, Xi'an , Shaanxi 710038, China.
ACS Appl Mater Interfaces. 2016 Apr 13;8(14):9014-21. doi: 10.1021/acsami.6b02662. Epub 2016 Apr 1.
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.
微小 RNA（miRNA）是一类参与包括致癌在内的各种生理过程的转录后基因调控因子，它们已成为肿瘤治疗学的潜在靶点。然而，反义寡核苷酸，称为抗-miRs，在体内拮抗 miRNA 功能的应用在很大程度上受到缺乏有效递药载体的阻碍。在这里，我们描述了聚酰胺胺（PAMAM）树枝状大分子和聚乙二醇（PEG）功能化纳米氧化石墨烯（NGO）缀合物（NGO-PEG-树枝状大分子）的开发，用于有效递送至非小细胞肺癌细胞中的抗-miR-21。为了监测抗-miR-21 递送至细胞和肿瘤内，我们还构建了一种包含三个与 miR-21 在报告基因 3'UTR 完全互补序列的可激活荧光素酶报告基因（Fluc-3xPS）。与裸树枝状大分子和 Lipofectamine 2000（Lipo2000）相比，NGO-PEG-树枝状大分子显示出相当低的细胞毒性和更高的转染效率。通过体外生物发光成像和 Western blot 分析，NGO-PEG-树枝状大分子有效地将抗-miR-21 递送至细胞质中，并导致荧光素酶强度和 PTEN 靶蛋白表达的上调呈剂量依赖性。此外，NGO-PEG-树枝状大分子转染抗-miR-21 导致细胞迁移和侵袭的抑制作用强于裸树枝状大分子或 Lipo2000 转染。通过 NGO-PEG-树枝状大分子静脉内递抗-miR-21 导致 Fluc-3xPS 报告基因移植瘤区域内的生物发光信号显著增加。这些结果表明，NGO-PEG-树枝状大分子可以作为一种有效的、有潜力的 RNA 寡核苷酸递药载体。此外，将 NGO-PEG-树枝状大分子与可激活的荧光素酶报告基因相结合的策略允许对递药过程进行图像引导监测，从而为 RNA 为基础的癌症治疗提供了新的见解。
