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山东大学学报(医学版) ›› 2014, Vol. 52 ›› Issue (2): 12-15.doi: 10.6040/j.issn.1671-7554.0.2013.525

• 基础医学 • 上一篇    下一篇

CHRNA5基因表达下调对肺癌细胞VEGF表达的影响

贾颖,祖珊珊,郏雁飞,肖东杰,汪运山,马晓丽   

  1. 山东大学附属济南市中心医院中心实验室, 济南 250013
  • 收稿日期:2013-08-28 出版日期:2014-02-10 发布日期:2014-02-10
  • 通讯作者: 马晓丽, E-mail:mxl7125@126.com
  • 基金资助:

    国家自然科学基金(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)

Effect of down-regulated CHRNA5 gene expression on VEGF expression of lung cancer

JIA Ying, ZU Shan-shan, JIA Yan-fei, XIAO Dong-jie, WANG Yun-shan, MA Xiao-li   

  1. Central Laboratory, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, China
  • Received:2013-08-28 Online:2014-02-10 Published:2014-02-10

摘要:

目的  体外研究非小细胞肺腺癌A549细胞中CHRNA5基因表达下调对尼古丁促肺癌细胞增殖及VEGF表达的影响。方法   实验分为0μmol/L尼古丁组、1μmol/L尼古丁组、转染阴性对照组(1μmol/L尼古丁+si-NC)和转染组(1μmol/L尼古丁+CHRNA5-siRNA)。设计合成编码α5-nAChR的CHRNA5基因siRNA片段,脂质体介导转染A549细胞,RT-PCR检测CHRNA5-siRNA转染后α5-nAChR和VEGF mRNA水平的表达,Western blotting检测转染后α5-nAChR蛋白水平的表达,ELISA法检测转染后细胞上清液中VEGF蛋白水平的表达,CCK-8方法   检测α5-nAChR下调对尼古丁促肺癌细胞增殖的影响。结果   CHRNA5-siRNA转染A549细胞后,转染组α5-nAChR和VEGF mRNA表达和蛋白表达量明显低于转染阴性对照组(P<0.05),且显著抑制尼古丁促肺癌细胞的增殖(P<0.05)。结论    α5-nAChR表达下调能抑制尼古丁促肺癌细胞的增殖,降低VEGF 的表达。α5-nAchR及其信号转导系统在吸烟相关性肺癌发生、发展中可能发挥重要作用,可作为肺癌预后预测指标和治疗靶点。

关键词: CHRNA5基因;&alpha, 5-尼古丁乙酰胆碱受体;血管内皮生长因子;尼古丁;肺肿瘤

Abstract:

Objective   To explore the influence of small interfering RNA (siRNA) down-regulating α5-nAChR expression on nicotine-induced proliferation of human lung cancer cells and expression of VEGF in vitro. Methods  The cells were divided into 0μmol/L nicotine group, 1μmol/L nicotine group, si-NC group (1μmol/L nicotine+si-NC) and siRNA group (1μmol/L nicotine+CHRNA5-siRNA). siRNA fragment of CHRNA5 encoding α5-nAChR was constructed and transfected into human lung cancer A549 cell line with lipofectamine 2000. After CHRNA5-siRNA transfection, expressions of α5-nAChR and VEGF in A549 cell were detected by RT-PCR, Western blotting and ELISA. The nicotine-induced A549 cell proliferation was detected by CCK-8. Results  After CHRNA5-siRNA transfection, expressions of α5-nAChR and VEGF on mRNA and protein levels were significantly decreased compared with the si-NC group (P<0.05), and nicotine-induced A549 cell proliferation was significantly inhibited (P<0.05). Conclusion  Down-regulation of α5-nAChR expression could inhibit nicotine-induced cell proliferation and decrease VEGF expression of lung cancer. α5-nAChR and its signal transduction system may play an important role in the development of smoking-related lung cancer and can be used as predictors of lung cancer prognosis and therapeutic targets.

Key words: CHRNA5 gene; &alpha, 5-nicotinic acetylcholine receptor; Vascular endothelial growth factor; Nicotine; Lung neoplasm

中图分类号: 

  • R73-3
[1] 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. 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[J]. 山东大学学报(医学版), 2013, 51(9): 8-12.
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