| 朱传东,王礼学,王国相,丁建勋,徐瀚峰,童金龙,郑勤.新型纳米金材料的合成及其对HepG2细胞放射增敏的影响[J].中华放射医学与防护杂志,2016,36(12):881-887 |
| 新型纳米金材料的合成及其对HepG2细胞放射增敏的影响 |
| Synthesis of novel gold nanoparticles and its radiosensitizing effect on HepG2 cells |
| 投稿时间：2016-06-30 |
| DOI：10.3760/cma.j.issn.0254-5098.2016.12.001 |
| 中文关键词: 纳米金 半乳糖酸 γ-H2AX 放射增敏 肝癌细胞 |
| 英文关键词:Gold nanoparticle Galactose γ-H2AX Radiosensitization Hepatocellular carcinoma cells |
| 基金项目:江苏省自然科学基金面上项目（BK20141084） |
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| 中文摘要: |
| 目的 合成新型纳米金复合物GAL-PEG-GNPs，探讨其体外放射增敏效应及增敏机制。方法 制备GAL-PEG-GNPs并进行表征。将HepG2细胞分为对照组、GNPs组和GAL-PEG-GNPs组。CCK-8法检测GAL-PEG-GNPs的细胞毒性，并计算IC50值；TEM和ICP-MS检测细胞对2种纳米材料的摄取；克隆形成实验检测放射增敏水平；流式细胞术分析细胞周期的变化；免疫蛋白印迹分析细胞凋亡相关蛋白变化；酶标仪检测细胞内过氧化氢酶（CAT）、超氧化物歧化酶（SOD）、总还原型谷胱甘肽（GSH）的表达水平。结果 GNPs和GAL-PEG-GNPs溶液的紫外可见吸收峰分别位于520和530 nm，GNPs和GAL-PEG-GNPs的直径分别为（22.6±2.12）和（32.0±1.41）nm。GAL-PEG-GNPs的细胞毒性与GNPs类似（P>0.05），IC50值分别为5.001和4.997 μg/ml。GAL-PEG-GNPs被HepG2细胞的摄取量高于GNPs。GNPs和GAL-PEG-GNPs的放射增敏比（SER）分别为1.46和1.95。细胞经过处理后，处于G2/M期的比例明显提高（t=14.20，P<0.05）。GNPs组和GAL-PEG-GNPs组Cytochrome C、Bax、Caspase-3、Caspase-9的表达水平较对照组明显提高，而Bcl-2的表达呈现降低趋势。GAL-PEG-GNPs组CAT、SOD、总GSH较对照组均明显减少（t=12.34、29.39、12.85，P<0.05）。结论 GAL-PEG-GNPs对HepG2细胞具有较好的放射增敏效应，增敏机制可能与GNPs诱导大量自由基产生，进而启动凋亡基因程序有关。 |
| 英文摘要: |
| Objective To synthesize novel gold nanoparticles of GAL-PEG-GNPs, study its radiation effect on hepatocellular carcinoma cells HepG2 cells in vitro, and investigate the underlying mechanisms. Methods GAL-PEG-GNPs were synthesized and characterized successfully. HepG2 cells were divided into three groups of control, GNPs and GAL-PEG-GNPs. The cytotoxicities of these compounds were tested by the CCK-8 assay and their IC50 values of HepG2 cells were calculated. Cell uptake of nanoparticles was detected by TEM and ICP-MS. The radiosensitization effect of nanoparticles was tested by the colony formation assay. Cell cycle distribution was detected by FCM. The expressions of CAT, SOD, and total GSH were detected with a microplate reader, and the expressions of apoptosis-related proteins were tested by Western blot. Results The GNPs and GAL-PEG-GNPs had absorption peaks at 520 and 530 nm, respectively, and their diameters were (22.6±2.12) and (32.0±1.41) nm detected by ICP-MS. The GAL-PEG-GNPs and GNPs had similar cytotoxicity profiles (P>0.05), while GAL-PEG-GNPs could be more effectively uptaken by HepG2 cells than GNPs. The sensitive enhancement ratio (SER) of GNPs and GAL-PEG-GNPs to HepG2 cells were 1.46和1.95, respectively. The percentage of cells at phase of G2/M in HepG2 population treated with GNP was higher than that of untreated cells (t=14.20, P<0.05). The protein expressions of Cytochrome C, Bax, Caspase-3, and Caspase-9 were up-regulated while Bcl-2 expression was down-regulated in the cells treated with GNPs/radiation or GAL-PEG-GNPs/radiation. The expressions of CAT, SOD and total GSH in the GNP treated groups were significantly decreased compared with the control group(t=12.34, 29.39, 12.85, P<0.05). Conclusions GAL-PEG-GNPs has obvious radiosensitization effect on HepG2 cells, which is related to the induction of cell apoptosis and the generation of free radicals. |
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