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Abstract：
Metformin (MET), an oral antidiabetic drug, was reported to possess promising anticancer effects. We hypothesized that MET encapsulation in unique nanospanlastics would enhance its anticancer potential against HEP-2 cells. Our results showed the successful fabrication of Nano-MET spanlastics (d = 232.10 ± 0.20 nm; PDI = 0.25 ± 0.11; zeta potential = (-) 44.50 ± 0.96; drug content = 99.90 ± 0.11 and entrapment efficiency = 88.01 ± 2.50%). MTT assay revealed the enhanced Nano-MET cytotoxicity over MET with a calculated IC50 of 50 μg/mL and > 500 μg/mL, respectively. Annexin V/PI apoptosis assay showed that Nano-MET significantly decreased the percentage of live cells from 95.49 to 93.70 compared to MET and increased the percentage of cells arrested in the G0/G1 phase by 8.38%. Moreover, Nano-MET downregulated BCL-2 and upregulated BAX protein levels by 1.57 and 1.88 folds, respectively. RT-qPCR revealed that Nano-MET caused a significant 13.75, 4.15, and 2.23-fold increase in caspase-3, -8, and - 9 levels as well as a 100 and 43.47-fold decrease in cyclin D1 and mTOR levels, respectively. The proliferation marker Ki67 immunofluorescent staining revealed a 3-fold decrease in positive cells in Nano-MET compared to the control. Utilizing the combined Pathway-Enrichment Analysis (PEA) and Reactome analysis indicated high enrichment of certain pathways including nucleotides metabolism, Nudix-type hydrolase enzymes, carbon dioxide hydration, hemostasis, and the innate immune system. In summary, our results confirm MET cytotoxicity enhancement by its encapsulation in nanospanlastics. We also highlight, using PEA, that MET can modulate multiple pathways implicated in carcinogenesis.
摘要：
二甲双胍(MET),一种口服抗糖尿病药物，据报道具有有希望的抗癌作用。我们假设在独特的纳米推拉剂中封装MET会增强其对HEP-2细胞的抗癌潜力。我们的结果表明，成功制造了Nano-MET痉挛药（d=232.10±0.20nm；PDI=0.25±0.11；zeta电位=（-）44.50±0.96；药物含量=99.90±0.11，包封率=88.01±2.50％）。MTT分析显示Nano-MET的细胞毒性比MET增强，计算的IC50为50μg/mL和>500μg/mL，分别。膜联蛋白V/PI凋亡实验显示，与MET相比，Nano-MET可将活细胞的百分比从95.49显着降低至93.70，并将停滞在G0/G1期的细胞百分比增加8.38％。此外，Nano-MET下调BCL-2，上调BAX蛋白水平1.57和1.88倍，分别。RT-qPCR显示，Nano-MET导致caspase-3，-8和-9水平显着增加13.75、4.15和2.23倍，细胞周期蛋白D1和mTOR水平降低100和43.47倍。分别。增殖标记Ki67免疫荧光染色显示，与对照相比，Nano-MET中的阳性细胞减少了3倍。利用联合途径富集分析（PEA）和Reactome分析显示某些途径的高度富集，包括核苷酸代谢，Nudix型水解酶,二氧化碳水合,止血,和先天免疫系统。总之,我们的结果证实了MET通过将其封装在纳米推拉剂中而增强了细胞毒性。我们还强调，使用PEA，MET可以调节与癌变有关的多种途径。
