香烟烟雾提取物对气道上皮细胞增殖和FIP表达的影响.docx

香烟烟雾提取物对气道上皮细胞增殖和FIP200表达的影响 【关键词】 气道上皮细胞 Effect of cigarette smoke extract on proliferation and cycle of human airway epithelial cell and its FIP200 expression 【Abstract】 AIM: To study the effect of cigarette smoke extract on the proliferation and cycle of human airway epithelial cell and its mechanism. METHODS: After airway epithelial cells were treated with CSE, the proliferation was measured with MTT and the distribution of different cell cycles was detected with flow cytometry. FAKfamily interacting protein of Mr 200 000 mRNA expression level, FAKfamily interacting protein of Mr 200 000expression level and the association of FIP200 with focal adhesion kinasewere analyzed respectively by reverse transcriptionpolymerase chain reaction, Western blot and immunoprecipitation. RESULTS: CSE inhibited the proliferation of human airway epithelial cells and arrested the epithelial cells in G1 phase of cell cycle. The number of epithelial cells in S and G2 phase dramatically decreased . CSE increased the expression level of FIP200 mRNA and FIP200 and the association of FIP200 with FAK. The effects of CSE on airway epithelial cells were all in a doseandtime dependent manner. The expression of FIP200 mRNA and FIP200 and the association of FIP200 with FAK were positively correlated to the increased number of epithelial cells in G1 phase and negatively correlated to the proliferation and the number of epithelial cells in S and G2 phase. CONCLUSION: CSE can inhibit the proliferation of human epithelial cells and the increased expression of FIP200 may be one of the factors responsible for it. 【Keywords】 CSE; airway epithelial cell; cell proliferation; FIP200 【摘要】 目的： 研究香烟烟雾提取物对气道上皮细胞增殖和细胞周期的影响及其作用机制. 方法： 利用MTT法研究CSE对气道上皮细胞增殖的影响，采用流式细胞术分析气道上皮细胞细胞周期的影响，以RTPCR和Western blot检测气道上皮细胞FIP200 mRNA和蛋白表达，以免疫共沉淀检测FIP200与局灶粘着斑激酶连接状况. 结果： CSE干预的气道上皮细胞MTT吸光度比对照组明显降低，G1期细胞增多，S和G2期细胞明显减少，FIP200 mRNA, FIP200的表达水平以及FIP200与FAK的连接均显著增高，且随CSE干预时间和CSE浓度而增大；气道上皮细胞FIP200表达水平和FIP200与FAK连接的增高均与G1