二羟环氧苯并芘诱导转化与翻译延长因子的关系

The relationship between BPDE-induced transformation and translation elongation factor 1 alpha 1 gene

目的探讨真核翻译延长因子1α1(eTEF1α1)基因在反式二羟环氧苯并芘致癌机制中所起的作用。方法采用逆转录聚合酶链反应(RT-PCR)方法扩增真核翻译延长因子1α1基因全长,插入pcDNATM3.1 D irectional TOPO表达载体,以脂质体转染介导的技术和G418细胞筛选法转染人支气管上皮细胞,构建转基因稳定表达的细胞株。用半定量RT-PCR方法分析转基因表达产物,对转基因细胞株,进行双层软琼脂试验以确定基因的恶性特性。结果成功扩增出eTEF1α1基因全长,构建出稳定表达真核翻译延长因子1α1基因的细胞株。稳定转染eTEF1α1基因的细胞株,能在双层软琼脂中形成克隆。结论真核翻译延长因子1α1基因与反式二羟环氧苯并芘的恶性转化有关。

Objective To investigate the role of eukaryotic translation elongation factor 1 alpha 1 （eTEFlal） gene in the carcinogenesis of anti-benzo（a） pyrene-trans-7,8-dihydrodiol-9,10-epoxide （anti-BPDE）. Methods A full-length of eTEFlal was amplified by reverse transcription-polymerase chain reaction （ RT-PCR）, inserted into pcDNATM 3.1 Directional TOPO expression vector, transfected into 16HBE ceils to construct stable transfected 16HBE cells with eTEFlal by using lipofectin and G418 selection protecols. The expression products of transfected genes were analyzed by semi-quantity RT-PCR. Semi-solid agar was applied to identify the malignancy features of the genes. Results The full-length of eTEFlal gene was successfully amplified, the stable 16HBE cell lines with eTEFlctl was constructed. The stable transfected cell line can form clones in semi-solid ager. Conclusion It was suggested that eTEF1αl could be associated with the malignant transformation induced by anti-BPDE.