生物合成纳米银的制备、表征及体外抗肿瘤活性评价

Omama E. Elshawy, E. Helmy, L. Rashed
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引用次数: 33

摘要

本研究以金黄色青霉(Penicillium aurantiogresium, IMI 89372)为主要材料合成银纳米粒子,研究其对MCF-7、MCT癌细胞和Vero(正常)细胞株在不同浓度(0.44 ~ 145 μg/ml)作用24 h后的细胞毒性。倒置光镜下观察细胞形态。此外,银纳米粒子(AgNPs)对MCF-7的放射增敏作用也通过评估细胞形态、MTT试验的细胞增殖、LDH活性和通过检查一些在癌变过程中改变的凋亡基因(包括caspase-3、Bax和Bcl-2)诱导凋亡来证明。同时估计Caspase-3活性。通过紫外可见光谱确定AgNPs的合成,并通过TEM、FT-IR和x射线分析(EDX、XRD)对其进行表征。通过直接电镜观察,生物合成的AgNPs为球形,尺寸为12.7 nm。生物合成AgNPs对MCF-7、MCT和Vero细胞系的细胞毒性表现出浓度依赖性,细胞形态发生不同程度的改变。结果表明,AgNPs对MCF-7具有高毒性,IC50值为10.5 μg/ml。辐照前处理MCF-7 (10.5 μg/ml)可通过增加细胞形态学改变、抑制细胞增殖、激活乳酸脱氢酶(LDH)和caspase-3导致细胞凋亡,从而改善辐照剂量(6 Gy)的效果,这一效应通过核DNA损伤增加、caspase 3和Bax基因上调、Bcl-2基因下调进一步得到证实。总之,本研究结果清楚地表明AgNPs具有剂量依赖性的细胞毒性,并证实AgNPs作为一种有效的放射增敏剂,可以增强γ辐射诱导的MCF-7乳腺癌细胞的杀伤。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Preparation, Characterization and in Vitro Evaluation of the Antitumor Activity of the Biologically Synthesized Silver Nanoparticles
This study was concentrated on the biosynthesis of silver nanoparticles from Penicillium aurantiogresium (IMI 89372) with a focus on its cytotoxicity in MCF-7 and MCT cancer cell lines as well as Vero (normal) cell line that was assessed by crystal violet assay after treatment with various concentrations (0.44 – 145 μg/ml) for 24 h. The cell morphology was examined by inverted light microscopy. Further, the radiosensitizing effect of silver nanoparticles (AgNPs) on MCF-7 was also demonstrated by assessing cell morphology, cell proliferation of MTT assay, LDH activity and induction of apoptosis through checking of some apoptotic genes that altered during carcinogenesis, including caspase-3, Bax and Bcl-2. Caspase-3 activity was also estimated. Synthesis of AgNPs was determined by UV-Visible spectrum and it was further characterized by TEM, FT-IR and X-Ray analysis (EDX, XRD). The biosynthesized AgNPs were spherical and of 12.7 nm in size as recorded by direct electron microscopy visualization. The biosynthesized AgNPs showed variation in cytotoxicity against MCF-7, MCT and Vero cell lines in a concentration dependant response with a varied degree of alteration in cell morphology. The result showed that AgNPs were highly toxic towards MCF-7 with IC50 value of 10.5 μg/ml. Treatment of MCF-7 (10.5 μg/ml) prior to irradiation improved the effect of irradiation dose (6 Gy) via increasing alteration of cell morphology, inhibition of cell proliferation, activation of the lactate dehydrogenase (LDH) and caspase-3 leading to induction of apoptosis which was further confirmed through increasing nuclear DNA damage and up regulation of caspase 3 and Bax genes and downrgulation of Bcl-2 genes. In conclusion, the present findings clearly indicated that AgNPs showed dose dependant cytotoxicity and verified that AgNPs acted as a potent radiosensitizer and could enhance gamma irradiation induced killing of MCF-7 breast cancer cells.
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