α5-烟碱型乙酰胆碱受体表达下调对肺癌细胞HIF-1α表达的影响  

Effects of dow n-regulated α5-nicotinic acetylcholine receptor expression on HIF-1α expression in lung cancer cells

作　　者：祖珊珊[1] 马晓丽[1,2] 郏雁飞[1,2] 赵云[3] 贾颖[1] 肖东杰[1,2] 汪运山[1,2] 

机构地区：[1]山东大学附属济南市中心医院中心实验室,济南250013 [2]山东省肿瘤防治靶向分子研究重点实验室,济南250013 [3]山东省警官总医院血液净化科,济南250002

出　　处：《山东大学学报（医学版）》2013年第9期8-12,共5页Journal of Shandong University:Health Sciences

基　　金：国家自然基金面上项目(81272588);山东省自然科学基金(ZR2012HM061);山东省高等学校科技计划(J11LC14);国家重点基础研究发展计划(973计划)(2012CB966503-4)

摘　　要：目的通过下调α5-烟碱型乙酰胆碱受体(α5-nAChR)的表达,体外研究尼古丁促肺癌细胞增殖过程中α5-nAChR对缺氧诱导因子(HIF-1α)表达的影响。方法以不同浓度尼古丁作用人肺腺癌细胞株A549,分别采用实时荧光定量PCR和Western blotting检测A549细胞中α5-nAChR、HIF-1αmRNA和蛋白的表达。MTT法、实时荧光定量PCR、Western blotting检测CHRNA5-siRNA转染后,尼古丁促A549细胞增殖的作用和HIF-1α表达的变化。结果尼古丁浓度为1μmol/L时,α5-nAChR和HIF-1α表达明显升高(P<0.05),二者表达呈尼古丁浓度依赖性。CHRNA5-siRNA转染显著抑制α5-nAChR、HIF-1α表达(P<0.05),尼古丁促A549细胞增殖的作用明显受到抑制(P<0.05)。结论α5-nAchR通过调节HIF-1α的表达,在尼古丁促肺癌细胞增殖中起作用,提示α5-nAChR/HIF-1α信号轴可能是吸烟相关性肺癌发生的重要分子机制之一。Objective To explore the influence of small interfering RNA （siRNA） down-regulated ct5-nicotinic acetyl- choline receptor （ct5-nAChR） expression on hypoxia-induced factor-1 ct （ HIF-1 α ） expression during the process of nico- tine-induced lung cancer A549 cell proliferation in vitro. Methods Expressions of ctS-nAChR and HIF-l~ at mRNA and protein levels in A549 cells were detected by FQ-PCR and Western blotting after treatment with various concentra- tions of nicotine. After CHRNAS-siRNA transfection, nicotine-induced A549 cell proliferation and expression of HIF- 1 oL were detected by MTT, FQ-PCR and Western blotting respectively. Results Expressions of ctS-nAChR and HIF- 1 cx were significantly enhanced with 1 txmol/L nicotine treatment compared with the other concentrations （ P 〈 0.05 ）, which presented a nicotine dose-dependent manner. After CHRNA5-siRNA transfection, the expressions of α5-nAChR mRNA and protein decreased obviously（ P 〈 0.05 ）. Nicotine-induced A549 cell proliferation was significantly inhibited （ P 〈 0.05）. Conclusion a5-nAchR plays an important role in nicotine-induced A549 cell proliferation via regulating HIF-I a expression, which suggests that α5-nAChR/HIF-1 oL axis may be one of the important molecular mechanisms for nicotine-related lung cancer.

关 键 词：CHRNA5基因 α5-烟碱型乙酰胆碱受体 缺氧诱导因子 尼古丁 肺肿瘤 

分 类 号：R733[医药卫生—肿瘤]