论文部分内容阅读
目的 克隆苯并 (a)芘代谢物二羟环氧苯并芘 (dihydroxyepoxybenzopyrene ,BPDE)诱导的人上皮细胞致癌相关差异表达cDNA序列。方法 以BPDE诱导恶性转化的人支气管上皮细胞株16HBE为模型 ,采用cDNA代表性差异分析方法 ,比较转化细胞及正常对照细胞间基因表达的差异 ,分离恶变细胞中差异表达的cDNA片段。分离的片断分别与pGEM T载体连接并转化入JM10 9细菌中 ,对质粒DNA进行测序。以BLASTN程序进行GenBank的同源性检索。结果 克隆的 13条cDNA序列中 ,有 5条为新基因序列 ,已在dbest数据库中注册 ,其登录号分别为BG35 46 91、BG35 46 92、BG35 46 93、BG35 46 94和BG35 46 95 ;8条cDNA有同源序列 ,分别与核糖体蛋白S2 3、MLN137、ACTN4、转移生长因子及G蛋白基因等有同源性。结论 克隆的 13条cDNA序列可能与BPDE诱发细胞恶性转化密切相关。
Objective To clone the differentially expressed cDNA sequence of human epithelial cells induced by dihydroxyepoxybenzopyrene (BPDE), a metabolite of benzo (a) pyrene. Methods 16HBE, a malignant transformed human bronchial epithelial cell line induced by BPDE, was used as a model. The difference of gene expression between transformed and normal control cells was compared by cDNA representative differential analysis, and the cDNA fragments differentially expressed in malignant cells were isolated. The isolated fragments were each ligated with the pGEM T vector and transformed into JM109 bacteria to sequence the plasmid DNA. Homology search of GenBank with BLASTN program. Results Five of the 13 cloned cDNA sequences were newly sequenced and have been registered in the dbest database with accession numbers of BG35 46 91, BG35 46 92, BG35 46 93, BG35 46 94 and BG35 46 95; 8 The cDNAs have homologous sequences and have homology with ribosomal protein S2 3, MLN137, ACTN4, metastatic growth factor and G protein gene, respectively. Conclusion The cloned 13 cDNA sequences may be closely related to BPDE-induced malignant transformation.