The toxicity of NaF on BmN cells and a comparative proteomics approach to identify protein expression changes in cells under NaF-stress Impact of NaF on BmN cells
The toxicity of NaF on BmN cells and a comparative proteomics approach to identify protein expression changes in cells under NaF-stress Impact of NaF on BmN cells
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DOI：
10.1016/j.jhazmat.2014.12.056
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发表时间：
2015-04-09
影响因子：
13.6
通讯作者：
Chen, Keping
中科院分区：
文献类型：
--
作者：
Chen, Liang;Chen, Huiqing;Chen, Keping
氟化物对生物体的发育产生负面影响，对人类健康和环境安全构成威胁。本研究以马里家蚕N细胞系（Bombyx mari N cell lines, BmN）为研究对象，探讨NaF对昆虫细胞的影响。我们发现高浓度NaF （>= 1 mM）培养8 h后，细胞形态发生了显著变化。NaF对连续培养72h的BmN的剂量-反应曲线显示，NaF对BmN的半数抑制浓度（IC50）为56.60 μ M， 100 μ M和300 μ M的NaF处理可诱导细胞凋亡和坏死。从BmN提取的全细胞二维电泳结果显示，与对照组相比，300 μ M NaF处理使32个蛋白上调，11个蛋白下调。我们通过MALDI-TOF MS鉴定了5种不同的蛋白，其中4种为首次鉴定，包括2种上调蛋白（线粒体醛脱氢酶ALDH2和禁止蛋白WPH）和2种下调蛋白（钙网蛋白前体CRT和DNA超卷曲因子SCF）。荧光定量PCR进一步证实了这些观察结果。综上所述，我们的数据表明这些靶蛋白可视为受NaF影响的靶标，也为研究NaF胁迫下的反应代谢途径提供了线索。(C) 2015 Elsevier B.V.版权所有
Fluorides negatively affect the development of organisms and are a threat to human health and environmental safety. In this study, Bombyx mari N cell line (BmN) were used to explore effects of NaF on insect cells. We found that 8 h (hrs) culture with high concentration of NaF (>= 1 mM) induced significantly morphological changes. Dose-response curves of 72 h continuously cultured BmN treated with NaF showed that the half inhibitory concentration (IC50) value was 56.60 mu M. Treatment of BmN with 100 and 300 mu M of NaF induced apoptosis and necrosis. 2-D electrophoresis of whole cell extracted from BmN showed that treatment with 300 mu M NaF up-regulated 32 proteins and down-regulated 11 proteins when compared with controls. We identified 5 different proteins by MALDI-TOF MS, and 4 of them were identified for the first time, including 2 up-regulated proteins (mitochondrial aldehyde dehydrogenase ALDH2 and prohibitin protein WPH) and 2 down-regulated proteins (calreticulin precursor CRT and DNA supercoiling factor SCF). These observations were further confirmed by fluorescence quantitative PCR. Together, our data suggest that these target proteins could be regarded as targets influenced by NaF and also provide clues for studies on the response metabolism pathway under NaF stress. (C) 2015 Elsevier B.V. All rights reserved.