State Key Laboratory of Elemento-Organic Chemistry and Department of Chemical Biology, National Engineering Research Center of Pesticide (Tianjin), College of Chemistry, Nankai University, 300071, Tianjin, China.
Chembiochem. 2022 Nov 4;23(21):e202200387. doi: 10.1002/cbic.202200387. Epub 2022 Sep 27.
Gene therapy offers an alternative and promising avenue to lung cancer treatment. Here, we used dibenzocyclooctyne (DBCO)-branched primers to construct a PTEN gene nanovector (NP-PTEN) through branch-PCR. NP-PTEN showed the nanoscale structure with biocompatible size (84.7±11.2 nm) and retained the improved serum stability compared to linear DNA. When transfected into NCI-H1299 cancer cells, NP-PTEN could overexpress PTEN protein to restore PTEN functions through the deactivation of PI3K-AKT-mTOR signaling pathway to inhibit cell proliferation and induce cell apoptosis. The apoptosis rate of NCI-H1299 cancer cells could reach up to 54.5 %±4.6 % when the transfection concentration of NP-PTEN was 4.0 μg/mL. In mice bearing NCI-H1299 tumor xenograft intratumorally administrated with NP-PTEN, the average tumor volume and tumor weight was separately reduced by 61.7 % and 63.9 %, respectively, compared with the PBS group on the 18 day of administration. The anticancer efficacy of NP-PTEN in NCI-H1299 tumor xenograft suggests the promising therapeutic potential of branch-PCR assembled PTEN gene nanovectors in lung cancer gene therapy and also provided more opportunities to introduce two or more tumor suppressor genes as an all-in-one gene nanovector for multiple gene-based cancer gene therapy.
基因治疗为肺癌治疗提供了一种有前途的替代方法。在这里，我们使用二苯并环辛炔（DBCO）分支引物通过分支 PCR 构建了 PTEN 基因纳米载体（NP-PTEN）。NP-PTEN 显示出纳米级结构，具有生物相容性尺寸（84.7±11.2nm），与线性 DNA 相比，保留了增强的血清稳定性。当转染到 NCI-H1299 癌细胞中时，NP-PTEN 可以过表达 PTEN 蛋白，通过失活 PI3K-AKT-mTOR 信号通路来恢复 PTEN 功能，从而抑制细胞增殖并诱导细胞凋亡。当 NP-PTEN 的转染浓度为 4.0μg/mL 时，NCI-H1299 癌细胞的凋亡率可高达 54.5±4.6%。在经 NP-PTEN 瘤内给药的荷 NCI-H1299 肿瘤异种移植小鼠中，与 PBS 组相比，给药 18 天后，平均肿瘤体积和肿瘤重量分别降低了 61.7%和 63.9%。NP-PTEN 在 NCI-H1299 肿瘤异种移植中的抗癌疗效表明，分支 PCR 组装的 PTEN 基因纳米载体在肺癌基因治疗中具有有前途的治疗潜力，也为引入两个或更多肿瘤抑制基因作为一种用于多种基于基因的癌症基因治疗的一体式基因纳米载体提供了更多机会。
