内源性大麻素联用顺铂对人肺腺癌A549细胞的诱导凋亡作用

Effects of AEA Combined with Cisplatin on Proliferation and Apoptosis of Human Lung Cancer A549 Cell Line

目的观察内源性大麻素(AEA)、顺铂(DDP)单用或联用对人肺癌A549细胞增殖抑制和诱导凋亡的作用。方法采用四甲基偶氮唑蓝(MTT)法检测AEA和DDP对肺癌A549细胞的增殖抑制作用,以流式细胞仪(FCM)PI单染检测AEA联用DDP对细胞周期的影响,以AnnexinV/PI双染法检测AEA联用DDP对细胞凋亡的影响。结果AEA对肺癌细胞增殖有明显的抑制作用,且呈剂量、时间依赖性。10,20μmol/L的AEA和1,2mg/L的DDP联合作用24,48,72h后,细胞增殖抑制率显著高于单用组;两药联用后诱导细胞凋亡的作用显著增强,AEA可使A549细胞阻滞于G2/M期。结论AEA及DDP对肺癌A549细胞的增殖具有显著的抑制作用,并可诱导细胞凋亡,在一定范围内呈量效关系,且两者联合应用具有相加或协同作用。

Objective To investigate the effect of anandamide（AEA）, cisplatin （DDP） and their synergistic effect on the growth inhibition ratio, apoptosis and cell cycle distribution of human lung adenocareinoma cell line A549. Methods To assess the sensitivity of A549 cells to AEA and DDP, A549 cells were exposed to increasing doses of AEA with or without DDP. MTT was employed to investigate prolifera- tion. Flow cytometry was used to investigate the apoptosis and cell cycle distribution of A549 cells affected by AEA, DDP and their combi- nation. Results Compared with either AEA or DDP alone, combining AEA at 10 or 20 p^mol/L with DDP at 1 or 2 mg/L respectively in- creased the growth inhibition rate of A549 cells （all P〈0.05） significantly. The apoptosis ratio increased according to the AEA and DDP concentration, and especially when DDP was combined with AEA. A549 cells showed G2/M stage arrest after AEA administration, and showed A549 cell S stage arrest with DDP administration（P 〈0. 01）. Conclusion Both AEA and DDP could enhance the growth inhibition and apoptosis of human lung cancer A549 cells in a concentration-dependent pattern. When they were combined, the effect could be enhanced in a synergistic or additive pattern.