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二羟环氧苯并芘诱发细胞恶变后基因和蛋白表达差异分析
Differential analysis of genes and protein expressions in malignantly transformed cells induced by trans-BPDE
【摘要】 目的研究苯并(a)芘代谢产物反式二羟环氧苯并芘(反式-BPDE)诱发人支气管上皮细胞系(16HBE)恶性转化的基因和蛋白表达的改变,探讨苯并(a)芘致癌的分子机制。方法应用含4 096条人类全长基因的cDNA芯片检测反式-BPDE诱发人支气管上皮细胞恶性转化的基因表达谱的变化;应用二维凝胶电泳技术和相应软件Im ageM aster2D 3.10分析反式-BPDE诱发人支气管上皮细胞恶性转化的蛋白质组变化,基质辅助激光解吸电离-飞行时间-质谱结合数据库检索鉴定部分表达有差异的蛋白斑点。结果BPDE诱发的恶性转化细胞与阴性对照细胞比较,cDNA芯片结果显示差异表达基因有143条,其中52条基因在BPDE诱发的恶性转化细胞中表达增高,91条基因在恶性转化细胞中表达降低;二维凝胶电泳显示有72个蛋白斑点出现表达差异,其中44个蛋白斑点在BPDE诱发的恶性转化细胞中表达升高,28个蛋白斑点在恶性转化细胞中表达降低,对其中7个表达明显差异的蛋白斑点的鉴定显示,原癌基因v-erb-b2同源3、锌指蛋白127、硫氧还原蛋白过氧化物酶2、K IAA0471蛋白在恶性转化细胞中高表达,而钙结合蛋白5、锌指蛋白A iolos、Kelch样蛋白3在恶性转化细胞中低表达。结论反式-BPDE诱发16HBE恶性转化的基因表达谱和蛋白表达谱发生了显著的变化,这些表达改变的基因和蛋白可能参与了BPDE诱发细胞恶变的过程。
【Abstract】 Objective To explore molecular mechanism of B(a)p carcinogenesis, and to study the alteration of gene and protein expressions in human bronchial epithelial malignantly transformed cells induced by benzo(a)pyrene metabolite trans-benzo(a)pyrene diol-epoxide (trans-BPDE). Methods The altered gene expression patterns in trans-BPDE-induced malignantly transformed cells were detected by cDNA microarray which representing 4 096 different genes. The differential protein expression patterns in malignantly transformed cells were analyzed by two dimensional electrophoresis (2-DE) and the ImageMaster 3.10 software. Matrix-assisted laser desorption/inoization-time of flight mass spectrometry (MALDI-TOF-MS) combined with database search was applied to identify the differentially expressed proteins. Results Compared with those of the negative control cells, 143 genes in BPDE-induced malignantly transformed cells were altered in expression, 52 of which were up-regulated and 91 were down-regulated; the levels of 72 proteins in malignantly transformed cells were changed, 44 of which were increased, while 28 decreased. And 7 proteins were identified by MALDI-TOF-MS, showing v-erb-b2 erythroblastic leukemia viral oncogene homolog 3, zinc finger protein 127, KIAA0471 protein (Fragment), and peroxiredoxin 2 were significantly up-regulated, while calcium-binding protein 5, zinc finger protein Aiolos, Kelch-like protein 3 were significantly down-regulated in malignantly transformed cells. Conclusions There are significant alterations of gene and protein profiles in malignantly transformed cells induced by trans-BPDE . The altered genes and proteins might be associated with B(a)p carcinogenesis.
【Key words】 Trans-BPDE; Human bronchial epithelial cells; Malignantly transformation; cDNA Microarray; Proteome;
- 【文献出处】 中国职业医学 ,Chinese Occupational Medicine , 编辑部邮箱 ,2005年05期
- 【分类号】R730.2
- 【被引频次】8
- 【下载频次】229