隐丹参酮抗肺腺癌作用的研究

Effect of cryptotanshinone on human lung adenocarcinoma cells

目的探讨隐丹参酮(CPT)对人肺腺癌细胞株A549和PC-9诱导凋亡的作用及其可能机制。方法不同浓度的CPT分别作用A549和PC-9细胞24和48h,CCK-8法检测细胞存活率,Annexin V-FITC/PI流式细胞术检测细胞凋亡,JC-1染色流式细胞术检测线粒体膜电位的变化,Western blot法检测凋亡相关蛋白cleaved Caspase-9、cleaved Caspase-3、聚腺苷酸二磷酸核糖转移酶(PARP)和信号转导及转录激活因子3(STAT3)信号通路相关蛋白磷酸化STAT3(p-STAT3)蛋白的表达水平。结果 CPT能够抑制A549和PC-9细胞的增殖,且呈剂量和时间依赖性。CPT作用于A549和PC-9细胞24h后,细胞凋亡率随CPT浓度增加呈增长趋势,而线粒体膜电位降低。CPT作用于A549和PC-9细胞后cleaved Caspase-9、cleaved Caspase-3和cleaved PARP蛋白表达水平增加,p-STAT3蛋白表达水平下降。结论 CPT对人肺腺癌细胞A549和PC-9有显著的增殖抑制作用和诱导凋亡效应,线粒体凋亡途径和STAT3信号通路可能参与了这一过程。

Objective To investigate the effect and related mechanism of cryptotanshinone(CPT)on human lung adenocarcinoma cells.Methods Human lung adenocarcinoma A549 and PC-9 cells were incubated with various concentrations of CPT for 24 and 48h,and cell viability was measured by CCK-8 assay.Cell apoptotic rate was detected by Annexin V-FITC/PI staining assay.Mitochondrial membrane potential(MMP)was evaluated by JC-1 staining assay.Levels of apoptosis related proteins and phosphorylation level of protein in key signaling pathways were examined by Western blot.Results CPT reduced the viability of A549 and PC-9 cells in a dose-and time-dependent manner.According to FCM analysis,the apoptotic rate increased with the increase of the concentration of CPT,and MMP also was decreased by CPT treatment.The protein expressions of cleaved Caspase-9,cleaved Caspase-3 and cleaved PARP were up-regulated,while the protein expression of p-STAT3 was down-regulated after CPT treatment.Conclusion CPT can inhibit the proliferation and induce apoptosis of human lung adenocarcinoma A549 and PC-9 cells,and mitochondrial apoptosis pathway and STAT3 signal pathway might be involved in the process.