【摘要】 目的探讨反式二羟环氧苯并芘(反式-BPDE)诱导转化(16HBE-T)与成瘤(16HBE-C)人支气管上皮细胞(16HBE)翻译延长因子1α1基因的表达变化。方法联合应用抑制性消减杂交、生物信息学分析以及半定量RT-PCR等方法,分别以16HBE-T和16HBE-C细胞cDNA为检测子,以正常16HBE细胞(16HBE-N)cDNA为驱动子,构建消减杂交文库,经2次杂交和2次PCR后,插入TA克隆载体克隆,经筛选、测序、序列分析后,设计特异引物,以β-肌动蛋白(β-actin)为内参照,进行半定量PCR。结果有9条差异表达片段与Genbank的翻译延长因子1α1的不同片段相同,半定量PCR表明,翻译延长因子1α1在16HBE-T和16HBE-C细胞中的表达水平分别为16HBE-N细胞的1.148倍和1.253倍,表明表达水平上调。结论翻译延长因子1α1可能与反式-BPDE的转化及成瘤有部分关系。更多还原

【Abstract】 Objective To investigate the expression changes of translation elongation factor 1α1 in anti-BPDE transformed and carcinoma 16 HBEs. Methods Suppression subtractive hybridization (SSH), bioinformatics and semi-quantitative RT-PCR were applied. The cDNA of anti-BPDE transformed and carcinoma 16 HBE cells were used as tester respectively, and the cDNA of normal 16 HBE was used as driver, the library of subtractive hybridization were profiled and inverted into TA cloning vector after two times of hybridization and two times of PCR. After the screening, sequencing and analysis of sequences, semi-quantitative RT-PCR was performed accompanying to the inner reference of β-actin. Results 9 differentially expressed fragments were consistent with translation elongation factor 1α1 in different regions in Genbank, and the expressions were up regulated in BPDE transformed and carcinoma 16 HBE cells. Conclusion Translation elongation factor 1α1 may be related to the transforming effect and carcinogenesis of anti-BPDE.更多还原