诺帝对恶性胶质瘤细胞株U87MG生物学行为的影响及差异表达蛋白质组分析  

Effects of nordy on biological behaviors of malignant glioma cell line U87MG and the analysis of differential expression proteome

作　　者：许建平[1] 刘宏[2] 卞修武[2] 陈剑鸿[2] 周向东[3] 吴玉章[4] 

机构地区：[1]第三军医大学西南医院病理学研究所第三军医大学新桥医院病理科,重庆400038 [2]第三军医大学西南医院病理学研究所,重庆400038 [3]第三军医大学西南医院病理学研究所基础部药学教研室,重庆400038 [4]第三军医大学西南医院免疫学研究所,重庆400038

出　　处：《中华病理学杂志》2007年第9期609-613,共5页Chinese Journal of Pathology

基　　金：国家863计划基金(20060102Z4108);重庆市科技重大攻关项目基金(CSTC-2005AA500T)

摘　　要：目的观察诺帝对人恶性胶质母细胞瘤细胞株 U87MG 及其移植瘤生物学行为的影响,分析和鉴定诺帝诱导 U87MG 细胞分化时高表达的差异蛋白质。方法采用自行研制的化合物诺帝(纯度为99.87%)诱导 U87MG 细胞分化,再用 U87MG 细胞原位移植瘤模型裸鼠腹腔注射诺帝,观察肿瘤生长情况和胶质纤维酸性蛋白(GFAP)表达改变。应用蛋白质双向电泳和 PDQuest7.1软件对比分析诺帝诱导分化前后表达的差异蛋白质,采用基质辅助激光解析离子化飞行时间质谱进行鉴定并分析其功能。结果诺帝能显著抑制 U87MG 细胞体外增殖和原位移植瘤的生长(P<0.01)并诱导其分化,U87MG 细胞体外分化后高表达的差异蛋白质包括增殖相关基因 A、交替拼接因子3、真核转录启动因子5A、丝切蛋白1、β半乳糖苷酶结合凝集素、甘油醛-3-磷酸脱氢酶、α烯醇化酶和一种未知蛋白。结论诺帝对人胶质母细胞瘤细胞株 U87MG 具有诱导分化的作用,其过程涉及多种蛋白质的表达改变,其中大部分蛋白质表达下调,这些蛋白质参与细胞增殖、代谢、分化、凋亡及基因转录的调控。Objective To explore effects of nordy on biological behaviors of human malignant glioblastoma cell line U87MG in vitro and transplanted tumor in vivo, and to identify the differential proteome upon Nordy induced differentiation. Methods Glioblastoma U87MG cells were induced to differentiate by synthetic lipoxygenase inhibitor, Nordy. The drug was also given via peritoneal injection to nude mice （27 mg/kg body weight） bearing orthotopic transplanted tumors of U87MG cells in the brain. The tumor volumes and GFAP expression were measured. Total proteins of U87MG cells after Nordy treatment were analysed by two-dimensional gel electrophoresis. PDQuest 7. 1 computer software was used to compare protein profiles of the treated cells with that of untreated control. Differentially expressed proteins were then selected and characterized by matrix assisted laser desorption ionization-time of flight-mass spectrometry. The functional aspects of these proteins were analyzed by bioinformatics. Results Nordy suppressed both the proliferation of U87MG cells in vitro and the tumor growth of orthotopic transplanted tumors in vivo （ P 〈 0.01 ）. The differentially expressed proteins induced by Nordy included proliferation-associated gene A, alternative splicing factor ASF-3, eukaryotic translation initiation factor 5A, coii31in 1 （non-muscle）, beta galactoside binding lectin, glyceraldehyde-3-phosphate dehydrogenase, enolase 1 and an unknown protein. Conclusions Nordy promotes the differentiation of glioblastoma cells, by which it may serve as a therapeutic agent. Various proteins identified during Nordy-induced differentiation are involved in the cell proliferation, metabolism, differentiation, apoptosis and gene transcription.

关 键 词：去甲二氢愈创木酸 神经胶质瘤 肿瘤移植 光谱分析 质量 细胞系 肿瘤 

分 类 号：R739.41[医药卫生—肿瘤]