| 叶武,黄勍栋,唐婷玉,叶芃,杜坚宗.自噬抑制剂3-甲基腺嘌呤促进荜茇酰胺的抗肿瘤作用[J].浙江中西医结合杂志,2019,29(11): |
| 自噬抑制剂3-甲基腺嘌呤促进荜茇酰胺的抗肿瘤作用 |
| Autophagy inhibitor 3-methyladenine promotes anti-tumor effect of piperlongumine |
| 投稿时间：2019-04-29 修订日期：2019-07-02 |
| DOI： |
| 中文关键词: 荜茇酰胺 肺癌 细胞自噬 3-甲基腺嘌呤 |
| 英文关键词:piperlongumine lung cancer autophagy 3-methyladenine |
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| 中文摘要: |
| 摘 要 目的 荜茇酰胺是一种中药单体，具有多种药理学作用。本研究旨在探讨自噬抑制剂3-甲基腺嘌呤对荜茇酰胺抗肿瘤作用的影响。方法 实验分为对照组：采用生理盐水处理A549细胞48h；荜茇酰胺组：3μM荜茇酰胺处理细胞48h，荜茇酰胺+ N-乙酰-L-半胱氨酸（NAC）组：3μM荜茇酰胺和10mM NAC处理细胞48h；3-甲基腺嘌呤+荜茇酰胺组：3μM荜茇酰胺和10mM NAC处理细胞48h。采用CCK-8法测定细胞活力，使用流式细胞仪测定细胞内活性氧（ROS）水平，蛋白质印迹法检测细胞自噬蛋白（LC3B）的表达。结果 3μM荜茇酰胺处理A549细胞后，细胞ROS水平显著升高至1.49±0.18(P<0.05)，细胞活力显著下降至60.1±10.2%（P<0.05），LC3B-II表达显著增加（P<0.05）。荜茇酰胺与10mM NAC合用后，细胞ROS水平为0.98±0.04，细胞活力为97.7+4.0%，LC3B-II表达下降至用药前水平（P<0.05）。自噬抑制剂3-甲基腺嘌呤处理肺癌细胞后，LC3B-II表达显著降低。3-甲基腺嘌呤+荜茇酰胺组A549细胞活力为24.8±1.3%，显著低于荜茇酰胺组（60.1±10.2%，P<0.05）。结论 荜茇酰胺通过ROS依赖途径促进细胞自噬。自噬抑制剂3-甲基腺嘌呤增强荜茇酰胺的抗肿瘤作用。 |
| 英文摘要: |
| ABSTRACT Objective Piperlongumine is a traditional Chinese medicine monomer with a variety of pharmacological effects. The aim of this study was to investigate the effects of autophagy inhibitor 3-methyladenine on the antitumor activity of piperlongumine. Methods The experiment was divided into the control group: A549 cells were treated with normal saline for 48 hours; piperlongumine group: 3 μM piperlongumine was used to treat cells for 48 h, piperlongumine+N-acetyl-L-cysteine (NAC) group: cells were treated with 3 μM piperlongumine and 10 mM NAC for 48 h; piperlongumine+3-methyladenine group: 3 μM piperlongumine and 10 mM 3-methyladenine were used to treat cells for 48 h. The cell viability of lung cancer cells was determined by CCK-8 assay, the level of reactive oxygen species (ROS) was determined by flow cytometry, and the expression of autophagy protein (LC3B) was detected by Western blotting. Results After treatment of A549 cells with 3μM piperlongumine, the ROS level increased significantly to 1.49±0.18（P<0.05）, the cell viability decreased significantly to 60.1±10.2%（P<0.05）, and the expression of LC3B-II increased significantly（P<0.05）. Following the combined treatment with piperlongumine and 10mM NAC, the intracellular ROS and cell viability returned to normal levels（P<0.05）, and the expression of LC3B-II decreased to the pre-dose level（P<0.05）. The viability of A549 cells in the 3-methyladenine combined with piperlongumine treatment group was 24.8±1.3%, which was significantly lower than that in the piperlongumine -treated group (60.1±10.2%, P<0.05). Conclusion Piperlongumine promotes cell autophagy through ROS-dependent pathways. The autophagy inhibitor 3-methyladenine enhances the anti-tumor effect of piperlongumine. |
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