Biopharmaceutical R&D Center, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou, China.
Biopharmagen Corp., Suzhou, China.
Sci Rep. 2019 Dec 11;9(1):18836. doi: 10.1038/s41598-019-55280-x.
An in vitro-transcribed RNA aptamer (trans-RA16) that targets non-small cell lung cancer (NSCLC) was previously identified through in vivo SELEX. Trans-RA16 can specifically target and inhibit human NCI-H460 cells in vitro and xenograft tumors in vivo. Here, in a follow-up study, we obtained a chemically-synthesized version of this RNA aptamer (syn-RA16) and a truncated form, and compared them to trans-RA16 for abilities to target and inhibit NCI-H460 cells. The syn-RA16, preferred for drug development, was by design to differ from trans-RA16 in the extents of RNA modifications by biotin, which may affect RA16's anti-tumor effects. We observed aptamer binding to NCI-H460 cells with K values of 24.75 ± 2.28 nM and 12.14 ± 1.46 nM for syn-RA16 and trans-RA16, respectively. Similar to trans-RA16, syn-RA16 was capable of inhibiting NCI-H460 cell viability in a dose-dependent manner. IC values were 118.4 nM (n = 4) for syn-RA16 and 105.7 nM (n = 4) for trans-RA16. Further studies using syn-RA16 demonstrated its internalization into NCI-H460 cells and inhibition of NCI-H460 cell growth. Moreover, in vivo imaging demonstrated the gradual accumulation of both syn-RA16 and trans-RA16 at the grafted tumor site, and qRT-PCR showed high retention of syn-RA16 in tumor tissues. In addition, a truncated fragment of trans-RA16 (S3) was identified, which exhibited binding affinity for NCI-H460 cells with a K value of 63.20 ± 0.91 nM and inhibited NCI-H460 cell growth by 39.32 ± 3.25% at 150 nM. These features of the syn-RA16 and S3 aptamers should facilitate the development of a novel diagnostic or treatment approach for NSCLC in clinical settings.
一种通过体内 SELEX 筛选出的针对非小细胞肺癌 (NSCLC) 的体外转录 RNA 适体 (trans-RA16)，可以在体外特异性靶向和抑制人 NCI-H460 细胞，并在体内抑制异种移植肿瘤。在此项后续研究中，我们获得了这种 RNA 适体的化学合成版本 (syn-RA16) 和一个截短形式，并将它们与 trans-RA16 进行比较，以评估它们靶向和抑制 NCI-H460 细胞的能力。syn-RA16 更适合药物开发，其设计在 RNA 修饰程度上与 trans-RA16 有所不同，修饰程度由生物素决定，这可能会影响 RA16 的抗肿瘤效果。我们观察到适体与 NCI-H460 细胞的结合，syn-RA16 的 K 值为 24.75±2.28 nM，trans-RA16 的 K 值为 12.14±1.46 nM。与 trans-RA16 相似，syn-RA16 能够以剂量依赖的方式抑制 NCI-H460 细胞的活力。syn-RA16 的 IC 值为 118.4 nM（n=4），trans-RA16 的 IC 值为 105.7 nM（n=4）。进一步使用 syn-RA16 的研究表明，它可以进入 NCI-H460 细胞并抑制其生长。此外，体内成像显示 syn-RA16 和 trans-RA16 逐渐在移植肿瘤部位积累，qRT-PCR 显示 syn-RA16 在肿瘤组织中有很高的保留率。此外，还鉴定出 trans-RA16 的一个截短片段 (S3)，其对 NCI-H460 细胞的结合亲和力的 K 值为 63.20±0.91 nM，在 150 nM 时抑制 NCI-H460 细胞生长 39.32±3.25%。syn-RA16 和 S3 适体的这些特征将有助于开发用于 NSCLC 的新型诊断或治疗方法。
