用余甘子合成的银纳米粒子对 Hep2 细胞系的抗增殖作用。
Antiproliferative effect of silver nanoparticles synthesized using amla on Hep2 cell line.
机构信息
Department of Biochemistry and Biotechnology, Annamalai University, Annamalai nagar-608 002, Tamilnadu, India.
出版信息
Asian Pac J Trop Med. 2013 Jan;6(1):1-10. doi: 10.1016/S1995-7645(12)60193-X.
OBJECTIVE
To synthesize silver nanoparticles by amla extract, screen the cytotoxic, oxidative stress and apoptotic effect of silver nanoparticles (AgNPs) on Hep2 cell line (laryngeal carcinoma cells) in vitro, and to compare the effect of Phyllanthus emblica (P. emblica) (amla) with AgNPs synthesized by amla and 5-FU.
METHODS
AgNPs was synthesized by P. emblica (aqueous extract) and nanoparticles were characterized UV-Vis spec, the presence of biomoloecules of amla capped in AgNPs was found by FT-IR analysis, shape and size were examined by SEM and DLS. Cytotoxicity of experimental drugs was tested to find IC(50) value. ROS generation in cells have been measured by DCFH-DA staining, AO-EtBr, Rhodamine-123 staining and DNA fragmentation were performed to assess apoptotic cell death, mitochondrial membrane potential and apoptotic DNA damage, respectively. Oxidative stress was analyzed by measuring lipid peroxides and antioxidants level to understand the cancer cell death by pro-oxidant mechanism.
RESULTS
PE-AgNPs was synthesized and confirmed through kinetic behavior of NPs. The shape of PE-AgNPs was spherical and cubic since it was agglomerated, and the nanoparticle surface was complicated. Average particle size distribution of PE-AgNPs was found to be 188 nm. Potent biomolecules of P. emblica such as polyphenols were capped with AgNPs and reduced its toxicity. In cytotoxicity assay the concentration in which the maximum number of cell death was 60 μg/mL and 50 μg/mL for P. emblica (alone) and AgNPs, respectively and IC(50) values were fixed as 30 μg/mL and 20 μg/mL. ROS generation, apoptotic morphological changes, mitochondrial depolarization, DNA damage and oxidative stress was observed as more in AgNPs treated cells than in P. emblica (30 μg/mL) (alone) treated cells and 5-FU treated cells gave similar result.
CONCLUSIONS
The results suggest that the AgNPs are capped with biomolecules of amla enhanced cytotoxicity in laryngeal cancer cells through oxidative stress and apoptotic function on Hep2 cancer cells.
目的
用余甘子提取物合成银纳米粒子，筛选体外对 Hep2 细胞系（喉癌细胞）的细胞毒性、氧化应激和凋亡作用，并比较余甘子（余甘子）与余甘子合成的银纳米粒子（AgNPs）和 5-FU 的作用。
方法
用余甘子（水提物）合成 AgNPs，通过紫外可见光谱对纳米粒子进行表征，通过傅里叶变换红外分析发现余甘子中生物分子的存在，通过扫描电子显微镜和动态光散射检查形状和大小。测定实验药物的细胞毒性，以找出 IC50 值。通过 DCFH-DA 染色、AO-EtBr 染色、罗丹明 123 染色来测量细胞内 ROS 的产生，分别进行凋亡细胞死亡、线粒体膜电位和凋亡 DNA 损伤的评估，通过测量脂质过氧化物和抗氧化剂水平来分析氧化应激，以了解通过促氧化剂机制导致癌细胞死亡。
结果
通过 NPs 的动力学行为合成并证实了 PE-AgNPs。PE-AgNPs 的形状为球形和立方体形，因为它发生了团聚，纳米粒子表面变得复杂。发现 PE-AgNPs 的平均粒径分布为 188nm。余甘子中的有效生物分子如多酚被 AgNPs 包裹，降低了其毒性。在细胞毒性试验中，最大细胞死亡浓度分别为 60μg/ml 和 50μg/ml，对于余甘子（单独）和 AgNPs，IC50 值分别为 30μg/ml 和 20μg/ml。与余甘子（30μg/ml）（单独）处理细胞相比，AgNPs 处理细胞中观察到 ROS 生成、凋亡形态变化、线粒体去极化、DNA 损伤和氧化应激增加，而 5-FU 处理细胞则产生类似的结果。
结论
结果表明，AgNPs 被余甘子的生物分子包裹，通过氧化应激和凋亡作用增强了喉癌细胞中对 Hep2 癌细胞的细胞毒性。
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