上海交通大学学报（医学版）

›› 2013, Vol. 33 ›› Issue (4): 421-.doi: 10.3969/j.issn.1674-8115.2013.04.008

• 论著（基础研究） • 上一篇    下一篇

低氧条件下RNA干扰阻断Notch3通路对人肺腺癌A549细胞增殖的影响

韩瑞超1,2, 范志文3, 周 敏1,3   

  1. 1.上海交通大学附属第六人民医院金山分院呼吸科, 上海 201500； 2.上海交通大学 医学院附属新华医院呼吸科, 上海 200092； 3.上海交通大学 医学院附属第三人民医院呼吸科, 上海 201900
  • 出版日期:2013-04-28 发布日期:2013-05-03
  • 作者简介:韩瑞超（1982—）, 男, 硕士生； 电子信箱: ruichaohan@gmail.com。
  • 基金资助:

    上海市金山区科学技术创新基金(2011-3-06) ；上海市教委科研创新项目（11YZ44）

Effects of blocking Notch3 pathway by siRNA under hypoxic condition on proliferation of human lung adenocarcinoma A549 cells

HAN Rui-chao1,2, FAN Zhi-wen3, ZHOU Min1,3   

  1. 1.Department of Respiratory Medicine, Jinshan Branch of the Sixth People´s Hospital, Shanghai Jiaotong University, Shanghai 201500, China； 2.Department of Respiratory Medicine, Xinhua hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200092, China； 3.Department of Respiratory Medicine, the Third People´s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 201900, China
  • Online:2013-04-28 Published:2013-05-03
  • Supported by:

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

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摘要：

目的 观察低氧条件下RNA干扰阻断Notch3对人肺腺癌A549细胞增殖的影响。方法 低氧条件下（1% O2）培养人肺腺癌A549细胞，以Notch3 siRNA转染A549细胞（Notch3 siRNA组），RT-PCR和Western blotting法分别检测A549细胞内Notch3及其信号通路下游基因HES1 mRNA和蛋白表达，MTT法检测转染24、48、72 h A549细胞的细胞活力；另设阴性对照组和空白对照组。结果 Notch3 siRNA转染A549细胞后，Notch3、HES1 mRNA和蛋白表达均显著降低（P<0.05）；Notch3 siRNA 转染后时间依赖性抑制A549细胞生长，Notch3 siRNA组转染48、72 h细胞活力与阴性对照组和空白对照组比较，差异有统计学意义（P<0.05）。结论 低氧条件下Notch3 siRNA转染人肺腺癌A549细胞可有效抑制Notch3及其下游靶基因HES1的表达，并抑制A549细胞生长。

关键词: Notch3, 肺腺癌, RNA干扰, 低氧

Abstract:

Objective To observe the effects of blocking Notch3 by siRNA under hypoxic condition on the proliferation of human lung adenocarcinoma A549 cells. Methods Human lung adenocarcinoma A549 cells were cultured under hypoxic condition (1% O2). After transfection with Notch3 siRNA (Notch3 siRNA group), RT-PCR and Western blotting were used to detect the expression of Notch3 and its downstream gene HES1 mRNA and protein in A549 cells respectively, and the cell viability 24 h, 48 h and 72 h after transfection was determined by MTT assay. Besides, negative control group and blank control group were established. Results After transfection with Notch3 siRNA, the expression of Notch3 and HES1 mRNA and protein in A549 cells was significantly decreased （P<0.05）. Transfection with Notch3 siRNA time-dependently inhibited the growth of A549 cells, and the cell viability 48 h and 72 h after transfection in Notch3 siRNA group was significantly different from those in negative control group and blank control group （P<0.05）. Conclusion Transfection with Notch3 siRNA under hypoxic condition may effectively inhibit the expression of Notch3 and its downstream gene HES1 in human lung adenocarcinoma A549 cells, and may suppress the growth of A549 cells.

Key words: Notch3, lung adenocarcinoma, siRNA, hypoxia