烟草凝聚物对人正常支气管上皮细胞中△Np63α表达的影响

摘要

[目的]探讨△Np63α与吸烟型肺癌发生的关系.[方法]用烟草凝聚物处理体外培养正常支气管上皮细胞HBE,并分别用Realtime PCR和Western blot检测其△Np63αmRNA和蛋白的表达.同时构建△Np63α的基因组启动子载体,利用荧光素酶分析烟草凝聚物对其转录活性的影响.[结果]烟草凝聚物处理后,HBE细胞中△Np63αmRNA和蛋白的表达明显增多,且具有剂量依赖性,以50 μg/mL浓度诱导效应最明显,mRNA和蛋白表达分别升高93％和311％.烟草凝聚物能够显著增强△Np63α启动子的转录活性,同样表现出剂量依赖性,50 μg/mL的诱导效应最明显,转录活性升高140％.[结论]烟草凝聚物能够通过增强△Np63α启动子的转录活性而诱导△Np63α的表达,说明△Np63α可能与吸烟导致的肺癌发生相关.%[Objective ] To study the correlation of ANp63α expression with cigarette smoking - induced lung cancer. [ Methods ] We cultured exposed human normal bronchial epithelial cells (HBE cells) to cigarette smoke extracts and detected the protein and mRNA levels of ANp63α with Western blot and realtime PCR respectively. We also cloned the ge-nomic promoter of ANp63α and determined its transcriptional activity after exposure to cigarette smoke extracts. [Results] The cigarette smoke extracts induced the expression of ANp63α both at protein and mRNA levels significantly in a dose dependent manner. The cigarette smoke extracts at 50μg/mL induced 93% and 311% increasing of mRNA and protein levels respectively. The transcriptional activity of the ANp63α promoter was also enhanced by cigarette smoke extracts in a dose dependent manner. The extracts at 50 μ.g/mL induced 140% increasing of transcriptional activity. [ Conclusion ] Cigarette smoke extracts could induce the expression of ANp63α by enhancing its transcriptional activity and this finding indicated that ANp63α might be involved in cigarette smoking - induced lung cancer.