苦参碱可能通过自噬抑制肺癌A549细胞的增殖

摘要

Objective:To investigate the inhibitory effect of matrine on the proliferation of lung cancer A549 cells and its effect on autophagy.Methods:A549 cells were cultured in vitro and treated with different concentrations of matrine.MTT assay was used to detect the cell proliferation inhibition rate.Inverted microscope were used to observe the cytomorphological changes of A549 cells.The suspension of varies concentration matrine to A549 cells were detec-ted by cell clone assay.The expression levels of Beclin1,p62 and LC3A/B were detected by Western blot.The chan-ges of autophagosome in A549 cells were observed under transmission electron microscope(TEM).Results:Matrine had a significant inhibitory effect on lung cancer A549 cells in a dose-dependent manner.Under inverted microsco-py,it was found that the lung cancer A549 cells appear shrinkage,smaller volume and visible granular particles of dif-ferent sizes after matrine treatment.With the increase of matrine concentration,cell colonies formed by lung cancer A549 cells gradually became smaller and less.The levels of Beclin1,p62 and LC3A/B were all increased with the in-crease of matrine concentration in a dose-dependent manner.It was observed by TEM that the autophagic vacuoles in A549 cells treated with matrine were significantly increased,and mitophagy was observed.Conclusion:Matrine may inhibit the proliferation of lung cancer A549 cells,induce autophagy and damage the autophagic pathways.Therefore, we speculate that matrine may inhibit the proliferation of lung cancer A549 cells through autophagy which disturbed the recycle of intracellular material and energy.%目的:探究苦参碱对肺癌A549细胞的增殖抑制作用及其对自噬的影响.方法:体外培养肺癌A549细胞,分别用不同浓度的苦参碱处理后,采用四甲基偶氮唑蓝(MTT)法检测细胞增殖抑制率;倒置显微镜下观察不同浓度苦参碱时A549细胞的形态变化;采用细胞克隆实验检测不同浓度的苦参碱对A549细胞的增殖抑制程度;Western blot检测自噬相关蛋白Beclin1、p62及LC3A/B的表达水平;透射电镜下观察经苦参碱处理后A549细胞内自噬泡的变化.结果:苦参碱对肺癌A549细胞有明显的抑制作用,并且呈剂量依赖性;倒置显微镜下发现经苦参碱处理后肺癌A549细胞皱缩、变小、胞内可见大小不一的颗粒状物质;随着苦参碱浓度的增加,肺癌A549细胞形成的克隆集落逐渐变小、变少;Western blot检测自噬相关蛋白Beclin1、p62及LC3A/B的水平均随着苦参碱浓度的增加而增加,并呈剂量依赖性;透射电镜下观察到苦参碱处理的肺癌A549细胞内自噬空泡明显增加,并且可观察到吞噬线粒体的自噬泡.结论:苦参碱可抑制肺癌A549细胞的增殖,并能诱导细胞内自噬的发生及损伤自噬降解通路,因此我们推测苦参碱可能通过自噬造成细胞内物质和能量的循环紊乱从而抑制肺癌A549细胞的增殖.