白花檵木对肺腺癌A549细胞增殖的抑制作用

Inhibitory Effect of Loropetalum Chinense on Proliferation of Lung Adenocarcinoma A549 Cells

目的探讨白花檵木粗提物对肺腺癌A549细胞体外增殖的影响。方法CCK-8法检测不同浓度白花檵木粗提物对A549细胞增殖的影响;集落形成实验检测白花檵木粗提物对A549细胞克隆生长能力的影响;流式细胞Annexin V-APC/PI双染法检测白花檵木粗提物诱导A549细胞凋亡的情况;免疫印迹法检测白花檵木粗提物处理后细胞凋亡相关蛋白Bax、Fas、Bcl-2、Caspase3以及活性Caspase3表达的影响。结果白花檵木粗提物可显著抑制A549细胞的生长增殖和集落形成,并呈时间及剂量依赖性,0.5 mg/ml白花檵木粗提物作用48 h后,A549细胞活力下降50%,40μg/ml白花檵木粗提物作用14天后,A549细胞集落形成能力被完全抑制;白花檵木粗提物作用A549细胞24 h后,呈现不同程度的凋亡,凋亡率随浓度的增加而升高;与对照组相比,凋亡抑制因子Bcl-2蛋白表达水平降低,Fas、Bax及活性Caspase3蛋白等凋亡促进因子表达水平升高。结论白花檵木粗提物体外对人肺腺癌细胞生长具有一定的抑制作用,其机制可能与激活线粒体及死亡受体凋亡通路有关。

Objective To investigate the effects of Loropetalum chinense extracts on the proliferation of lung adenocarcinoma A549 cells cultured in vitro.Methods CCK-8 assay was performed to evaluate the effect of Loropetalum chinense extracts on the proliferation of A549 cells,and the clonal growth ability of A549 cells was determined by clone formation assay.Flow cytometry Annexin V-APC/PI double staining was used to measure the apoptosis of A549 cells.Western blot was used to measure the expressions of apoptosisrelated proteins Bax,Fas,Bcl-2 Caspase3 and cleaved-Caspase 3.Results Loropetalum chinense extracts significantly inhibited the proliferation and colony formation of A549 cells in a time-and dose-dependent manner.The viability of A549 cells decreased by 50%after 48h treatment of 0.5mg/ml extracts,and 100%inhibition of colony formation rate achieved when the cells were treated with 40μg/ml extracts for 14 days.When A549 cells were treated with Loropetalum chinense extracts for 24h,apoptotic rates increased in a dose-dependent manner.Compared with the control group,the protein levels of Fas,Bax,Caspase3 and cleaved-Caspase3 were up-regulated,while Bcl-2 was down-regulated.Conclusion Loropetalum chinense extracts inhibit the growth of human lung adenocarcinoma cells in vitro,and the mechanisms may be related to the activation of mitochondrial and death receptor apoptosis pathway.