【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。更多还原

【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.更多还原