| 何 聪,陈清勇,王 剑.姜黄素下调mTOR诱导人肺癌A549细胞自噬的研究[J].肿瘤学杂志,2014,20(5):363-368. |
| 姜黄素下调mTOR诱导人肺癌A549细胞自噬的研究 |
| Curcumin Inducing Autophagy in Lung Cancer A549 Cells Through Down-regulation of mTOR |
| 投稿时间：2014-01-15 |
| DOI：10.11735/j.issn.1671-170X.2014.05.B003 |
|
 |
| 中文关键词: 姜黄素 自噬 微管相关蛋白轻链 哺乳动物雷帕霉素靶蛋白 肺肿瘤 |
| 英文关键词:curcumin autophagy microtubule associated protein light chain(LC3) mammals rapamycin target protein(mTOR) lung neoplasms |
| 基金项目:浙江省中医药科学研究基金计划（2011ZA093） |
|
| 摘要点击次数: 2148 |
| 全文下载次数: 753 |
| 中文摘要: |
| 摘 要：［目的］ 研究姜黄素诱导人肺癌A549细胞自噬现象及与哺乳动物雷帕霉素靶蛋白(mTOR)的关系。［方法］采用CCK-8法检测姜黄素对A549细胞增殖的抑制作用；采用吖啶橙染色(AO) 及转染GFP-LC3质粒荧光显微镜下观察姜黄素处理后A549细胞的自噬现象；采用Western blot法检测微管相关蛋白轻链Ⅱ(LC3Ⅱ)、微管相关蛋白轻链Ⅰ(LC3Ⅰ)、mTOR表达的变化。［结果］ 姜黄素对人肺癌A549细胞增殖有明显抑制作用，且成明显的剂量—时间依赖关系。AO染色结果显示，与对照组相比，姜黄素40μmol/L组及Rapa组细胞内酸性滤泡染成亮红色荧光比例增多；与姜黄素40μmol/L组相比，姜黄素40μmol/L+自噬抑制剂3-甲基腺嘌呤(3-MA)组及3-MA组亮红色荧光比例下降。细胞转染GFP-LC3后姜黄素40μmol/L组和Rapa组胞浆内点状的绿色荧光斑点明显；与姜黄素40μmol/L组相比，姜黄素40μmol/L+3-MA组及3-MA组胞浆内点状的绿色荧光斑点明显比例有所下降。Western blot法检测结果显示，与对照组相比，姜黄素40μmol/L组作用24h后，LC3-Ⅱ/LC3-Ⅰ表达显著升高（P＜0.05），姜黄素40μmol/L + 3-MA组LC3Ⅱ/LC3-Ⅰ表达低于姜黄素40μmol/L组（P＜0.05）。姜黄素40μmol/L组mTOR表达显著下降（P＜0.01），姜黄素40μmol/L+3-MA组mTOR表达高于姜黄素40μmol/L组（P＜0.05）。 ［结论］ 姜黄素能够明显抑制A549细胞增殖并诱导A549细胞发生自噬，同时还能够明显抑制mTOR蛋白的表达水平，其机理可能与mTOR的信号通路有关。 |
| 英文摘要: |
| Abstract：［Purpose］ To study the effect of curcumin inducing autophagy in human lung cancer A549 cells and the association with mTOR. ［Methods］ The inhibition of curcumin on A549 cells were detected by CCK-8 method. Autophagy was observed by the fluorescence microscope through acridine orange staining and the GFP-LC3 plasmid transfecting.Expression of microtubule associated protein light chain Ⅱ (LC3Ⅱ)，microtubule associated protein light chain Ⅰ(LC3Ⅰ)，mammals rapamycin target protein(mTOR) changes were evaluated by Western blot method. ［Results］ The proliferation of A549 lung cancer cells were significantly inhibited by curcumin in a dose-time dependent manner. AO staining showed that，compared with control group，the intracellular acid dye follicular bright red fluorescence ratio were increased in the curcumin 40μmol/L group and Rapa group. Compared with curcumin 40μmol/Lgroup，red fluorescence ratio were decreased in curcumin 40μmol/L combined with autophagy inhibitor 3-methyl adenine(3-MA) group and 3-MA group. Intracytoplasmic GFP-LC3 dotted green fluorescence spots were significantly increased in the curcumin 40μmol/L and Rapa group，and decreased in curcumin 40μmol/L+3-MA group and 3-MA group compared with 40μmol/L curcumin group. Compared with control group，LC3-Ⅱ/LC3-Ⅰexpression significantly increased in curcumin 40μmol/L group(P<0.05). LC3-Ⅱ/LC3-Ⅰexpression decreased in curcumin 40μmol/L+3-MA group compared with curcumin 40μmol/L group(P<0.05). mTOR expression was significantly decreased in curcumin 40μmol/L group(P<0.01)，while it was higher in curcumin 40μmol/L + 3-MA group(P<0.05).［Conclusion］ The proliferation of A549 cells could be significantly inhibited by curcumin，that may involve in cell autophagy，and mTOR may play an important role in this process. |
|
在线阅读
查看全文 查看/发表评论 下载PDF阅读器 |