Zughaibi Torki A, Jabir Nasimudeen R, Khan Azhar U, Khan Mohd Shahnawaz, Tabrez Shams
King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia.
Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.
Cell Biochem Funct. 2023 Dec;41(8):1174-1187. doi: 10.1002/cbf.3850. Epub 2023 Sep 10.
Cu O is the least explored copper oxide, and its nanoformulation is anticipated to have important therapeutic potential especially against cancer. The current study aimed to biosynthesize Cu O nanoparticles (NPs) using an aqueous extract of pumpkin seeds and evaluate its antiproliferative efficacy against cervical cells after screening on different cancer cell lines. The obtained NPs were characterized by different spectroscopic analyses, such as UV-vis, thermogravimetric, energy dispersive X-ray, and Fourier-transform infrared spectroscopy (FTIR). In addition, high-resolution transmission electron microscopes (HR-TEM) were used to observe the morphology of the biosynthesized NPs. The UV-vis spectra showed a peak at around 332 nm, confirming the formation of Cu O NPs. Moreover, FTIR and TAG analyses identified the presence of various bioactive phytoconstituents that might have worked as capping and stabilization agents and comparative stable NPs at very high temperatures, respectively. The HR-TEM data showed the spherical shape of Cu O NPs in the range of 100 nm. The Cu O NPs was screened on three different cancer cell lines viz., Hela, MDA-MB-231, and HCT-116 using cytotoxicity (MTT) reduction assay. In addition, Vero was taken as a normal epithelial (control) cell. The high responsive cell line in terms of least IC was further assessed for its anticancer potential using a battery of biological tests, including morphological alterations, induction of apoptosis/ROS generation, regulation of mitochondrial membrane potential (MMP), and suppression of cell adhesion/migration. Vero cells (control) showed a slight decline in % cell viability even at the highest tested Cu O NPs concentration. However, all the studied cancer cells viz., MDA-MB-231, HCT 116, and HeLa cells showed a dose-dependent decline in cell viability after the treatment with Cu O NPs with a calculated IC value of 10, 11, and 7.2 µg/mL, respectively. Based on the above data, Hela cells were chosen for further studies, that showed induction of apoptosis from 3.5 to 9-folds by three different staining techniques acridine orange/ethidium bromide (AO/EB), 4',6-diamidino-2-phenylindole (DAPI), and propidium iodide (PI). The enhanced production of reactive oxygen species (>3.5-fold), modulation in MMP, and suppression of cell adhesion/migration were observed in the cells treated with Cu O NPs. The current study obtained the significant antiproliferative potential of Cu O NPs against the cervical cancer cell line, which needs to be confirmed further in a suitable in vivo model. Based on our results, we also recommend the green-based, eco-friendly, and cost-effective alternative method for synthesizing novel nanoformulation.
氧化铜是研究最少的铜氧化物，其纳米制剂有望具有重要的治疗潜力，尤其是对抗癌症。当前的研究旨在使用南瓜籽水提取物生物合成氧化铜纳米颗粒（NPs），并在对不同癌细胞系进行筛选后，评估其对宫颈细胞的抗增殖功效。通过不同的光谱分析对获得的纳米颗粒进行表征，如紫外可见光谱、热重分析、能量色散X射线光谱和傅里叶变换红外光谱（FTIR）。此外，使用高分辨率透射电子显微镜（HR-TEM）观察生物合成纳米颗粒的形态。紫外可见光谱在332nm左右出现一个峰值，证实了氧化铜纳米颗粒的形成。此外，FTIR和热重分析分别鉴定出存在各种可能作为封端和稳定剂的生物活性植物成分，以及在非常高的温度下相对稳定的纳米颗粒。HR-TEM数据显示氧化铜纳米颗粒呈球形，粒径在100nm范围内。使用细胞毒性（MTT）还原试验在三种不同的癌细胞系，即Hela、MDA-MB-231和HCT-116上筛选氧化铜纳米颗粒。此外，将Vero细胞作为正常上皮（对照）细胞。对于IC值最低的高反应性细胞系，使用一系列生物学试验进一步评估其抗癌潜力，包括形态学改变、凋亡/活性氧生成诱导、线粒体膜电位（MMP）调节以及细胞粘附/迁移抑制。Vero细胞（对照）即使在最高测试的氧化铜纳米颗粒浓度下，细胞活力百分比也仅略有下降。然而，所有研究的癌细胞，即MDA-MB-2,31、HCT 116和HeLa细胞在用氧化铜纳米颗粒处理后，细胞活力均呈剂量依赖性下降，计算得到的IC值分别为10、11和7.2μg/mL。基于上述数据，选择HeLa细胞进行进一步研究，通过吖啶橙/溴化乙锭（AO/EB）、4',6-二脒基-2-苯基吲哚（DAPI）和碘化丙啶（PI）三种不同的染色技术，显示凋亡诱导倍数从3.5倍到9倍。在用氧化铜纳米颗粒处理的细胞中观察到活性氧的产生增加（>3.5倍）、MMP的调节以及细胞粘附/迁移的抑制。当前的研究获得了氧化铜纳米颗粒对宫颈癌细胞系的显著抗增殖潜力，这需要在合适的体内模型中进一步证实。基于我们的结果，我们还推荐这种基于绿色、环保且经济高效的替代方法来合成新型纳米制剂。
