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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。
Objective To investigate the expression of the translational elongation factor 1α1 gene in 16HBE-C and 16HBE-induced trans-BPDE induced transplantion in 16HBE-C human bronchial epithelial cells (16HBE). Methods Combined 16HBE-T and 16HBE-C cDNAs with 16HBE-C cDNA and 16HBE-N cDNA as the driving force, we used suppression subtractive hybridization, bioinformatics analysis and semi-quantitative RT- The subtractive hybridization library was constructed. After two hybridizations and two PCRs, the cloned TA cloning vector was inserted. After screening, sequencing and sequence analysis, specific primers were designed and β-actin was used as internal reference Semi-quantitative PCR. Results Nine different expression fragments were identical to the different fragments of translational elongation factor 1α1 in Genbank. Semi-quantitative PCR showed that the expression level of translation elongation factor 1α1 in 16HBE-T and 16HBE-C cells was 1.148 times higher than that in 16HBE-N cells And 1.253-fold, indicating that the expression level is up-regulated. Conclusion The translation elongation factor 1α1 may be partly related to the trans-BPDE transformation and tumorigenesis.