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| 人恶性胶质瘤细胞SHG-44经诺帝诱导分化后形态学改变及差异表达蛋白的质谱分析 |
| 许建平,卞修武,陈意生,吴玉章,蒋雪峰,XUJian-ping,BIANXiu-wu,CHENYi-sheng,WUYu-zhang,JIANGXue-feng |
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| 摘要: |
| 目的:观察人恶性胶质瘤细胞SHG-44经去甲二氢愈创木酸类似物--诺帝诱导分化后形态学的改变,并分析差异表达蛋白.方法:分别用100、200 μmol/L诺帝诱导SHG-44细胞分化,观察处理后24、48、72 h细胞形态学的改变,并与未受刺激的空白对照组相比较;提取200μmol/L诺帝处理后的SHG 44细胞以及空白对照组细胞总蛋白进行双向电泳,用PDqest7.1软件比较两者间的差异表达蛋白,离子飞行时间质谱仪分析高丰度表达的差异蛋白.结果:200 μmol/L诺帝处理后SHG-44细胞的形态学改变较100 μmol/L更为明显;处理72 h时细胞分化最为明显.与空白对照组SHG-44细胞相比,经200 μmol/L诺帝诱导后,双向电泳发现了23个差异蛋白点,其中21个蛋白点表达下调,2个蛋白点表达上调.质谱分析高丰度表达的差异蛋白分别为:未知蛋白、增殖相关基因A、解链蛋白Up1、交替拼接因子ASF-3、cofiln 1、真核转录启动因子5A、β半乳糖苷酶结合凝集素、Pi类谷光苷肽-S-转移酶.结论:诺帝能诱导人恶性胶质瘤细胞SHG-44分化,并呈一定的时效及量-效依赖性,分化后的差异蛋白涉及到细胞增殖、分化、凋亡及基因转录调控等多个方面. |
| 关键词: 去甲二氢愈创木酸、神经胶质瘤、细胞分化、细胞学、光谱分析,质量 |
| DOI：10.3724/SP.J.1008.2006.00378 |
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| 基金项目:国家“863”计划引导项目（2002AA001010）,重庆市科技攻关项目 |
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| Morphological changes and differential protein spectrum of human malignant glioma cell SHG-44 after treated with Nordy,an analog of Nordihydroguaiaretic acid |
| 许建平,卞修武,陈意生,吴玉章,蒋雪峰,XU Jian-ping,BIAN Xiu-wu,CHEN Yi-sheng,WU Yu-zhang,JIANG Xue-feng |
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| Abstract: |
| Objective：To observe the morphological changes and analyze the differential protein spectrum of human malignant glioma cells SHG-44 after treated with Nordy （Chinese patent number：ZL02133700.4）, an analog of Nordihydroguaiaretic acid. Methods： The differentiation of SHG-44 cells was induced by 100μmol/L or 200 μmol/L Nordy; the morphological changes of cells were observed 24, 48 and 72 h after Nordy treatment and the findings were compared with those of the control group （received no treatment） . The total proteins were extracted from SHG-44 cells treated with 200 μmol/L Nordy for 72 h and cells in control group, then were subjected to two-dimensional gel electrophoresis. PDquest 7. 1 software was employed to compare the protein expression differences. The highly expressed differential proteins were identified by matrix-assisted laser desorption/ ionization-time of flight-mass spectrometry （MALDI-TOF-MS）. Results： The morphological changes of SHG-44 cells treated with 200 μmol/L Nordy were more obvious than those treated with 100 μmol/L Nordy, and the most obvious differentiation was found in the cells treated for 72 h. Compared with those of control group, 23 differential protein spots were identified by the two-dimensional electrophoresis, including 21 down-regulated ones and 2 up regulated ones. MALDI-TOF-MS showed that the highly expressed proteins were： an unknown protein, proliferation-associated gene A, Upl, alternative splicing factor ASF-3, cofilinl（non-muscle）, eukaryotic translation initiation factor 5A, beta galactoside binding lectin, and glutathione-S-transferase Pi. Conclusion.. Nordy can induce differentiation of human malignant glioma cells SHCr-44 in a time-effect and dose-effect dependent manner. The Nordy-induced differential proteins may function in multiple aspects such as cell proliferation, apoptosis and gene transcription |
| Key words: nordihydroguaiaretic acid glioma cell differentiation cytology spectrum analysis, mass |
