PK-PD Tox & Formulation Section, Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India.
PK-PD Tox & Formulation Section, Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India.
Int J Pharm. 2024 Jan 5;649:123599. doi: 10.1016/j.ijpharm.2023.123599. Epub 2023 Nov 21.
Gemcitabine (GEM) is an important chemotherapeutic agent used alone or in combination with other anticancer agents for the treatment of various solid tumors. In this study, the potential of a dietary supplement, α-tocopherol succinate (TOS) was investigated in combination with GEM by utilizing human serum albumin-based nanoparticles (HSA NPs). The developed nanoparticles were characterized using DLS, SEM and FTIR and evaluated in a panel of cell lines to inspect cytotoxic efficacy. The ratio metric selected combination of the NPs was further investigated in human pancreatic cancer cell line (MIA PaCa-2 cells) to assess the cellular death mechanism via a myriad of biochemical and bio-analytical assays including nuclear morphometric analysis by DAPI staining, ROS generation, MMP loss, intracellular calcium release, in vitro clonogenic assay, cell migration assay, cell cycle analysis, immunocytochemical staining followed by western blotting, Annexin V-FITC and cellular uptake studies. The desolvation-crosslinking method was used to prepare the NPs. The average size of TOS-HSA NPs and GEM-HSA NPs was found to be 189.47 ± 5 nm and 143.42 ± 7.4 nm, respectively. In combination, the developed nanoparticles exhibited synergism by enhancing cytotoxicity in a fixed molar ratio. The selected combination also significantly triggered ROS generation and mitochondrial destabilization, alleviated cell migration potential and clonogenic cell survival in MIA PaCa-2 cells. Further, cell cycle analysis, Annexin-V FITC assay and caspase-3 activation, up regulation of Bax and down regulation of Bcl-2 protein confirmed the occurrence of apoptotic event coupled with the G0/G1 phase arrest. Nanocarriers based this combination also offered approximately 14-folds dose reduction of GEM. Overall, the combined administration of TOS-HSA NPs and GEM-HSA NPs showed synergistic cytotoxicity accompanied with dose reduction of the gemcitabine. These encouraging findings could have implication in designing micronutrient based-combination therapy with gemcitabine and demands further investigation.
吉西他滨（GEM）是一种重要的化疗药物，单独或与其他抗癌药物联合用于治疗各种实体瘤。在这项研究中，利用人血清白蛋白纳米粒（HSA NPs）研究了膳食补充剂α-生育酚琥珀酸酯（TOS）与 GEM 的联合应用。采用 DLS、SEM 和 FTIR 对所开发的纳米粒进行了表征，并在一系列细胞系中进行了细胞毒性评价。进一步在人胰腺癌细胞系（MIA PaCa-2 细胞）中研究了比例选择的 NPs 组合，通过 DAPI 染色核形态计量分析、ROS 生成、MMP 损失、细胞内钙释放、体外集落形成试验、细胞迁移试验、细胞周期分析、免疫细胞化学染色和 Western blot 分析、Annexin V-FITC 和细胞摄取研究等多种生化和生物分析方法评估细胞死亡机制。采用去溶剂交联法制备 NPs。发现 TOS-HSA NPs 和 GEM-HSA NPs 的平均粒径分别为 189.47±5nm 和 143.42±7.4nm。在固定摩尔比下，联合使用时，所开发的纳米粒表现出协同作用，增强了细胞毒性。所选组合还显著触发了 ROS 生成和线粒体失稳，减轻了 MIA PaCa-2 细胞的迁移潜力和集落形成细胞存活。此外，细胞周期分析、Annexin-V FITC 测定和 caspase-3 激活、Bax 上调和 Bcl-2 蛋白下调证实了与 G0/G1 期阻滞相关的凋亡事件的发生。基于这种组合的纳米载体还使吉西他滨的剂量减少了约 14 倍。总的来说，TOS-HSA NPs 和 GEM-HSA NPs 的联合给药表现出协同细胞毒性，并减少了吉西他滨的剂量。这些令人鼓舞的发现可能对设计基于微量营养素的吉西他滨联合治疗具有重要意义，需要进一步研究。
