上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (8): 1024-.doi: 10.3969/j.issn.1674-8115.2012.08.014

• 论著（基础研究） • 上一篇    下一篇

γ分泌酶抑制剂阻断Notch信号通路对肺腺癌细胞A549增殖的影响

周 敏1,2, 范志文1, 韩瑞超1   

  1. 1.上海交通大学 医学院附属第三人民医院呼吸科, 上海 201900； 2.上海市第六人民医院金山分院呼吸科, 上海 201500
  • 出版日期:2012-08-28 发布日期:2012-08-29
  • 作者简介:周 敏（1967—）, 女, 副主任医师, 硕士；电子信箱: zhoumin1209@163.com。
  • 基金资助:

    上海市教育委员会科研创新项目（11YZ44）和上海市金山区科学技术创新基金(2011-3-06)

Influence of blocking Notch signaling pathway by γ-secretase inhibitor on proliferation of lung adenocarcinoma A549 cells

ZHOU Min1,2, FAN Zhi-wen1, HAN Rui-chao1   

  1. 1.Department of Respiratory Medicine, the Third People´s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 201900, China；2.Department of Respiratory Medicine, Jinshan Branch of the Sixth People´s Hospital, Shanghai Jiaotong University, Shanghai 201500, China
  • Online:2012-08-28 Published:2012-08-29
  • Supported by:

    Shanghai Education Committee Foundation, 11YZ44；Shanghai Jinshan District Science and Technology Innovation Foundation, 2011-3-06

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摘要：

目的 观察γ分泌酶抑制剂MW167阻断Notch信号对肺腺癌细胞A549增殖的影响。方法 正常氧条件下培养肺腺癌细胞A549，通过Western blotting及RT-PCR方法检测Notch受体各亚型的表达。在正常氧条件下，不同浓度（5、10、20 μmol/L）MW167处理肺腺癌细胞A549（加药组），48 h后采用Western blotting方法检测Notch1蛋白表达的变化，采用RT-PCR方法检测Notch信号通路下游基因HES1mRNA表达的变化，应用MTT法检测MW167对细胞增殖的影响；每个浓度设二甲基亚砜对照组。结果 Notch1～4各亚型在A549细胞均有表达。5、10、20 μmol/L MW167处理肺腺癌A549细胞48 h后，各加药组与对照组相比，Notch1蛋白表达升高（P<0.05），而HES1 mRNA表达下降（P<0.05），随MW167浓度升高，对HES1 mRNA表达的抑制率逐渐升高（r=0.927，P<0.01）。MTT法检测结果显示各加药组吸光度高于对照组（P<0.05），且随MW167浓度升高，细胞增殖率相应升高（r=0.904，P<0.01）。结论 在正常氧条件下，MW167可阻断Notch信号通路，对A549细胞起促增殖作用；Notch1信号通路可能在A549细胞中起一定抑癌作用。

关键词: γ分泌酶抑制剂, Notch信号, 肺腺癌

Abstract:

Objective To observe the influence of blocking Notch signaling pathway by γ-secretase inhibitor MW167 on the proliferation of lung adenocarcinoma A549 cells. Methods Lung adenocarcinoma A549 cells were cultured in vitro under normoxic condition, and the expression of Notch receptor subtypes was detected by Western blotting and RT-PCR. Under normoxic condition, lung adenocarcinoma A549 cells were treated with different concentrations (5, 10 and 20 μmol/L) of MW167 (treatment groups). Forty-eight hours after treatment, the expression of Notch1 protein was determined by Western blotting, the expression of Notch signaling pathway downstream gene HES1mRNA was detected by RT-PCR, and the influence of MW167 on cell proliferation was determined by MTT. Besides, DMSO control groups of corresponding concentrations were established. Results Notch1-4 subtypes were expressed in A549 cells. Compared with control groups, the expression of Notch1 protein was higher (P<0.05) and that of HES1 mRNA was lower (P<0.05) in treatment groups 48 h after treatment, and the inhibition rate of HES1 mRNA expression increased with MW167 concentrations (r=0.927, P<0.01). The absorbance of each treatment group was higher than that of each control group (P<0.05), and the cell proliferation rates increased with MW167 concentrations (r=0.904, P<0.01). Conclusion Under normoxic condition, MW167 may block Notch signaling pathway and promote proliferation of A549 cells. Notch1 signaling pathway may play a tumor suppressing role in A549 cells.

Key words: γ-secretase inhibitor, Notch signal, lung adenocarcinomaNotch