Department of Genetics and Advanced Medical Technology, Medical Biotechnology Research Center, Faculty of Medicine, AJA University of Medical Sciences, Tehran 1411718541, Iran.
Department of Biotechnology, Alzahra University, Tehran 1993893973, Iran.
Molecules. 2022 Aug 24;27(17):5418. doi: 10.3390/molecules27175418.
A PEGylated niosomal formulation of cyclophosphamide (Nio-Cyclo-PEG) was prepared using a central composite design and characterized in terms of drug loading, size distribution, and average size. The stability of formulations was also studied at different conditions. In vitro cytotoxicity of drug delivery formulations was assessed on gastric cancer cells using MTT assay. The mechanism of cytotoxicity was studied at the transcriptional level by real-time PCR on Caspase3, Caspase9, CyclinD, CyclinE, MMP-2, and MMP-9 genes, while apoptosis was investigated with flow cytometry. The anti-metastatic property was evaluated using the scratch method. Propidium iodide staining was used to study the cell cycle. The results indicated that the as-designed nanocarrier exhibited a controlled drug release pattern with improved nanoparticle stability. It was found that the living cancer cells treated with Nio-Cyclo-PEG showed a significant decrease in number when compared with the niosomal carrier without PEG (Nio-Cyclo) and free drug (Cyclo). Moreover, the drug-loaded nanocarrier induced planned death (apoptosis) in the cancer cells through the regulation of Caspase3, Caspase9, CyclinD, CyclinE, MMP-9, and MMP-2 gene expression, indicating that the Nio-Cyclo-PEG formulation could significantly inhibit the cell cycle at the sub G1 phase as well as prevent the migration of cancer cells. In conclusion, Nio-Cyclo-PEG as developed in this study could serve as an active-targeting drug delivery nanocarriers for gastric cancer therapy with high efficacy and minimal side effects on healthy tissues/cells.
聚乙二醇化的脂质体环磷酰胺（Nio-Cyclo-PEG）制剂采用中心复合设计制备，并在药物载量、粒径分布和平均粒径方面进行了表征。还研究了制剂在不同条件下的稳定性。采用 MTT 法测定药物传递制剂对胃癌细胞的体外细胞毒性。通过实时 PCR 研究 Caspase3、Caspase9、CyclinD、CyclinE、MMP-2 和 MMP-9 基因的转录水平研究细胞毒性机制，同时通过流式细胞术研究细胞凋亡。采用划痕法评价抗转移特性。碘化丙啶染色研究细胞周期。结果表明，所设计的纳米载体表现出可控的药物释放模式，纳米颗粒稳定性得到提高。与未聚乙二醇化的脂质体（Nio-Cyclo）和游离药物（Cyclo）相比，用 Nio-Cyclo-PEG 处理的活癌细胞数量明显减少。此外，载药纳米载体通过调节 Caspase3、Caspase9、CyclinD、CyclinE、MMP-9 和 MMP-2 基因的表达，诱导癌细胞程序性死亡（凋亡），表明 Nio-Cyclo-PEG 制剂可显著抑制细胞周期在亚 G1 期，并防止癌细胞迁移。总之，本研究开发的 Nio-Cyclo-PEG 可作为一种主动靶向的胃癌治疗药物传递纳米载体，具有高效和对健康组织/细胞最小的副作用。
