白藜芦醇对非小细胞肺癌细胞自噬和凋亡的影响

目的 探讨白藜芦醇（resveratrol，Res）对人非小细胞肺癌A549细胞自噬和凋亡的影响。方法 采用50 μmol/L白藜芦醇作用于非小细胞肺癌A549细胞（Res组），对照组细胞用2% FBS的DMEM培养，培养24 h后采用倒置显微镜观察两组A549细胞形态的变化，激光扫描共聚焦显微镜观察细胞自噬小体的形成，Western blot检测自噬相关蛋白Beclin1、LC3Ⅱ/Ⅰ、p62和凋亡相关蛋白Bcl-2、Bax、活化caspase-3的表达。结果 A549细胞培养24 h后， 倒置显微镜下可见Res组细胞形状较对照组变圆，细胞排列稀疏；激光扫描共聚焦显微镜下可观察到Res组A549细胞胞浆内大量自噬体形成。Western blot检测结果显示，与对照组比较，Res组A549细胞自噬相关蛋白Beclin 1、LC3 Ⅱ/Ⅰ表达升高（0.151±0.032 vs 0.093±0.013，P=0.011；3.644±0.122 vs 1.903±0.054，P＜0.001），p62蛋白表达下降（0.032±0.002 vs 0.061±0.005，P=0.021）；促凋亡相关蛋白Bax、活化caspase-3表达升高（0.633±0.061 vs 0.423±0.053，P=0.011；0.154±0.004 vs 0.111±0.011，P=0.002），而抑制凋亡蛋白Bcl-2表达下降（2.437±0.055 vs 3.503±0.138，P＜0.001）。结论 白藜芦醇可通过促进非小细胞肺癌A549细胞过度自噬而诱导细胞凋亡。

Effect of resveratrol on autophagy and apoptosis of non-small cell lung cancer cells

Objective To investigate the effects of resveratrol （Res） on autophagy and apoptosis of non-small cell lung cancer A549 cells. Methods A549 cells were divided into control group and resveratrol group （Res group）. Res group cells were treated with 50 μmol/L resveratrol，and the control group cells were cultured with DMEM containing 2% FBS. The morphological changes were observed by inverted microscopy. The formation of autophagosomes was observed by laser scanning confocoal. The expression of the autophagy-related proteins Beclin1，LC3Ⅱ/Ⅰ，p62 and apoptosis-related proteins Bcl-2，Bax，Active caspase-3 were detected by Western blot. Results After 24 h culture，the cell in the Res group were more round and sparsely arranged than those in the control group. A large number of autophagy bodies formed in the cytoplasm. Western blot showed that Beclin1 and and LC3Ⅱ/Ⅰ expression were increased in A549 cells in the Res group than in the control group（0.151±0.032 vs 0.093±0.013，P=0.011；3.644±0.122 vs 1.903±0.054，P＜0.001），but p62 protein expression was reduced（0.032±0.002 vs 0.061±0.005，P=0.021） in the Res group. The expression of pro-apoptotic protein Bax and activated caspase-3 were increased in the Res group（0.633±0.061 vs 0.423±0.053，P=0.011；0.154±0.004 vs 0.111±0.011，P=0.002）. However，the expression of anti-apoptotic protein Bcl-2 was reduced in the Res group（2.437±0.055 vs 3.503±0.138，P＜0.001）. Conclusion Resveratrol induces non-small cell lung cancer A549 cells apoptosis by promoting excessive autophagy.