上海交通大学学报（医学版）

›› 2013, Vol. 33 ›› Issue (1): 25-.doi: 10.3969/j.issn.1674-8115.2013.01.005

• 论著（基础研究） • 上一篇    下一篇

细胞自噬对吉非替尼抑制肺癌细胞增殖的影响

徐志红1, 高蓓莉2, 胡家安1   

  1. 上海交通大学 医学院附属瑞金医院 1.老年病科, 2.呼吸科, 上海 200025
  • 出版日期:2013-01-28 发布日期:2013-02-06
  • 通讯作者: 胡家安, 电子信箱: jahu_rj@yahoo.com.cn。
  • 作者简介:徐志红（1971—）, 女, 博士, 副主任医师；电子信箱: zhihongx@hotmail.com。

Effects of autophagy on growth inhibition of Gefitinib in non-small cell lung cancer cell lines

XU Zhi-hong1, GAO Bei-li2, HU Jia-an1   

  1. 1.Department of Geriatrics, 2.Department of Respirology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2013-01-28 Published:2013-02-06

PDF (PC)

1374

摘要：

目的 探讨细胞自噬对吉非替尼（Gefitinib）抑制非小细胞肺癌（NSCLC）细胞增殖的影响。方法 采用Western blotting和LC3抗体检测自噬标志物LC3Ⅱ的含量，判断Gefitinib是否诱导NSCLC Calu6和H460细胞自噬。采用增强绿色荧光蛋白（EGFP）-LC3b质粒转染NSCLC细胞系Calu6和H460，再用40 μmol/L Gefitinib处理细胞48 h，观察绿色荧光蛋白（GFP）的分布情况。分别采用细胞自噬诱导剂依维莫司（Everolimus）、Gefitinib和Gefitinib联合Everolimus处理H460细胞，药物处理10 d后用结晶紫染色，显微镜下统计克隆数，计算不同药物刺激后细胞克隆形成数和药物对H460细胞的抑制率。结果 Western blotting检测显示，Gefitinib 40 μmol/L处理细胞48 h后，明显增加LC3Ⅱ的积累，绿色荧光呈散点状分布。经Everolimus、Gefitinib和Gefitinib联合Everolimus处理后，H460细胞克隆形成数差异有统计学意义（P<0.05），其中Gefitinib联合Everolimus处理对H460细胞的抑制率最强，两者之间有协同作用。结论 Gefitinib能够诱导NSCLC细胞自噬，并且增加细胞自噬能增强Gefitinib对肺癌细胞增殖的抑制率。

关键词: 非小细胞肺癌, 吉非替尼, 细胞自噬

Abstract:

Objective To investigate the effects of autophagy on growth inhibition of Gefitinib in non-small cell lung cancer (NSCLC) cell lines. Methods The content of autophagy marker LC3II was detected by Western blotting and LC3 antibody, and whether Gefitinib could induce autophagy in NSCLC Calu6 and H460 cells were determined. H460 and Calu6 cells were transfected with enhanced green fluorescence protein (EGFP)-LC3b plasmid, and were treated with 40 μmol/L Gefitinib for 48 h respectively. The distribution of green fluorescence protein (GFP) was observed. In addition, H460 cells were treated with Everolimus, Gefitinib, and Everolimus combined with Gefitinib respectively. Ten days later, cells were stained with crystal violet, cell clonies were counted under microscope, and growth inhibition rates were calculated. Results Western blotting revealed that LC3Ⅱ was significantly accumulated after treatment with 40 μmol/L Gefitinib for 48 h, and green fluorescence was dispersedly distributed. There were significant differences in numbers of clony formation among H460 cells treated with Everolimus, Gefitinib, and Everolimus combined with Gefitinib (P<0.05), and the growth inhibition rate of Everolimus combined with Gefitinib was the highest. Conclusion Gefitinib induces autophagy in NSCLC cells, and autophagy induction enhances growth inhibition of Gefitinib on NSCLC cells.

Key words: non-small lung cancer, Gefitinib, autophagy