二羟环氧苯并芘致肺癌相关基因筛选

Screening of lung tumor susceptibility gene caused by benzo(a) pyrene dihydrodiol epoxide

目的筛选与二羟环氧苯并芘,(BPDE)引发肺癌相关基因的DNA片段,为进一步探讨BPDE致癌机制提供研究依据。方法应用扩增片段长度多态性(AFLP)结合免疫磁珠分离等技术对与BPDE相互作用的DNA片段进行富集、扩增,经聚丙烯酰胺凝胶电泳显示差异片段,并对其回收、克隆、测序及同源相似性比对分析。结果成功回收了差异片段,经测序得到其碱基序列,基因同源相似性比对分析发现,有8条序列与已知基因同源,另有1条序列未找到同源序列,将该序列向GenBank提交注册,GenBalak已接受并给予登录号为:EF176681。结论AFLP结合免疫磁珠分离等技术可以用于化学致癌BPDE肿瘤易感基因的筛选,所得9条基因片段可能与BPDE引发肺癌有关。

Objective To screen tumor susceptibility gene of lung cancer induced by benzo[a]pyrene dihydrodiol epoxide（BPDE） interaction with mice lung DNA and settle a foundation for exploring carcinogenesis mechanism of BPDE.MethodsAmplified fragment length polymorohism（AFLP） combination with immunomagnetic beads isolation were used to enrich and amplify fragments of BPDE interaction with DNA.Differential fragments were displayed by polyacrylamide gel electrophoresis and silver nitrate staining.The differential fragments were sequenced and compared with known genes in genbank.Results 9 differential bands were successfully collectad,cloned and sequenced,and one fragment could not be found in the known genes,and then was submitted to genbank（accession number：EF 176681）.Conclusion AFLP combination with immunomagnetic beads isolation technique can be used to screen tumor susceptibility gene of chemaicl carcinogens BPDE,the 9 sequences may be relevant to lung cancer induced by BPDE.