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目的肺癌位居我国居民癌症发病率和死亡率的首位,其中非小细胞癌(non-small cell lung cancer,NSCLC)约占80%,筛选高效低毒的抗癌药物尤为迫切。本研究拟探讨姜黄素对NSCLC细胞的可能作用机制。方法用不同浓度的姜黄素(0、10、20、30μmol/L)或活性氧清除剂(CAT和NAC)加姜黄素处理肺癌细胞A549和SPC-A1,采用流式细胞术检测细胞周期、细胞凋亡率、活性氧(reactive oxygen species,ROS)水平和线粒体膜电位的变化,蛋白质印迹法检测自噬相关蛋白LC3、P62和细胞凋亡相关蛋白PARP、Caspase-3和Caspase-9的表达变化。结果姜黄素抑制非小细胞肺癌细胞株A549和SPC-A1增殖及克隆形成,主要将细胞阻滞在G_2/M期,0、10、20和30μmol/L姜黄素处理的A549细胞G_2/M期细胞百分比分别为(12.67±2.52)%、(22.67±2.52)%、(27.00±2.01)%和(42.33±4.04)%,SPC-A1细胞G_2/M期细胞百分比分别为(9.33±2.52)%、(18.33±1.53)%、(20.67±2.52)%和(30.67±1.53)%。0、10、20和30μmol/L姜黄素处理A549细胞凋亡率分别为(4.40±1.02)%、(7.31±1.52)%、(9.32±1.08)%和(13.97±1.98)%,P<0.05;SPC-A1细胞凋亡率分别为(4.38±1.22)%、(5.98±0.75)%、(9.42±1.25)%和(16.13±3.09)%,P<0.05。姜黄素导致ROS水平增高、线粒体膜电位降低和线粒体自噬的发生,并且呈剂量依赖性,而应用ROS清除剂可以减弱以上药物作用。相关信号转导通路蛋白表达与以上细胞生物行为改变表现一致。结论姜黄素通过ROS途径诱导NSCLC细胞发生线粒体自噬,是一种有潜力的抗癌药物。
The purpose of lung cancer ranks first in the incidence and mortality of cancer in our country residents, of which about 80% of non-small cell lung cancer (NSCLC), the screening of highly effective and low toxicity of anticancer drugs is particularly urgent. This study was to explore the possible mechanism of curcumin on NSCLC cells. Methods The lung cancer cells A549 and SPC-A1 were treated with different concentration of curcumin (0, 10, 20, 30μmol / L) and active oxygen scavengers (CAT and NAC) plus curcumin. The cell cycle was detected by flow cytometry. (ROS) and mitochondrial membrane potential were detected by Western blotting. The expression of autophagy-related proteins LC3, P62 and PARP, Caspase-3 and Caspase-9 were detected by Western blotting . Results Curcumin inhibited the proliferation and clonality of A549 and SPC-A1 cells in non-small cell lung cancer cell lines. The cells were mainly arrested in G 2 / M phase. The G 2 / M phase of A549 cells treated with 0, 10, 20 and 30 μmol / L curcumin The percentages of cells in the SPC-A1 cells were (12.67 ± 2.52)%, (22.67 ± 2.52)%, (27.00 ± 2.01)% and (42.33 ± 4.04)%, respectively , (18.33 ± 1.53)%, (20.67 ± 2.52)% and (30.67 ± 1.53)%, respectively. The apoptotic rates of A549 cells treated with 0,10,20 and 30μmol / L curcumin were (4.40 ± 1.02)%, (7.31 ± 1.52)%, (9.32 ± 1.08)% and (13.97 ± 1.98)%, respectively ; The apoptotic rates of SPC-A1 cells were (4.38 ± 1.22)%, (5.98 ± 0.75)%, (9.42 ± 1.25)% and (16.13 ± 3.09)% respectively, P <0.05. Curcumin resulted in increased ROS levels, decreased mitochondrial membrane potential and mitophagy, and in a dose-dependent manner. However, the use of ROS scavenger attenuated the above effects. The expression of the relevant signal transduction pathways is consistent with the above changes in cellular biological behavior. Conclusion Curcumin induces mitochondrial autophagy in NSCLC cells through the ROS pathway and is a promising anticancer drug.