摘要
目的探讨异黄腐酚(isoxanthohumol,IN)与顺铂(cisplatin,DDP)联合用药对肺癌DDP耐药A549/DDP细胞的协同抑制作用,并阐明其可能的分子机制。方法以CCK-8法检测IN、DDP以及两药联用对A549/DDP细胞增殖的影响,并通过Isobologram分析两药联用的协同作用;将A549/DDP细胞分为空白组(未加任何药物处理)、IN组(20μmol·L^(-1) IN)、DDP组(6 mg·L^(-1) DDP)、IN+DDP联合用药组(20μmol·L^(-1) IN+6 mg·L^(-1) DDP),流式细胞术检测细胞周期分布和细胞凋亡率;Western blot法检测各组细胞中耐药相关蛋白P-gp、LRP、MRP和PI3K/AKt通路中PI3K和p-AKt蛋白表达。结果与DDP组比较,IN与DDP联合用药可以明显降低DDP对A549/DDP细胞的IC50;Isobologram分析结果也表明二者联用对A549/DDP细胞具有协同抑制作用;与空白组、IN组、DDP组3组比较,IN+DDP联合用药组明显阻滞细胞周期在G0/G1期、增加A549/DDP细胞凋亡率(P<0.01)、降低A549/DDP细胞耐药蛋白P-gp、LRP和MRP的表达水平(P<0.01)及明显下调PI3K和p-AKt蛋白表达水平(P<0.01)。结论IN与DDP两药联用可协同抑制A549/DDP细胞增殖、阻滞细胞周期和促进细胞凋亡,其机制可能与是通过抑制耐药蛋白P-gp、LRP和MRP的表达,及抑制PI3K/AKt信号通路的活性,从而增强DDP对耐药细胞的作用有关。
Aim To investigate the synergistic inhibitory effect of isoxanthohumol(IN)and cisplatin(DDP)on lung cancer DDP-resistant A549/DDP cells,and to clarify its possible molecular mechanism.Methods The CCK-8 method was used to detect the effects of IN,DDP and the combination of the two drugs on the proliferation of A549/DDP cells,and the synergy of the two drugs was analyzed by Isobologram.The A549/DDP cells were divided into a blank group(without any Drug treatment),IN group(20μmol·L^(-1) IN),DDP group(6 mg·L^(-1) DDP),IN+DDP combination group(20μmol·L^(-1) IN+6 mg·L^(-1) DDP).Flow cytometry was used to detect cell cycle distribution and cell apoptotic rate.Western blot was used to detect the expression of PI3K and p-AKt proteins in the resistance-related proteins P-gp,LRP,MRP and PI3K/AKt pathway in each group of cells.Results Compared with DDP group,the combination of IN and DDP could significantly reduce the IC50 of DDP on A549/DDP cells.Isobologram analysis results also showed that the combination of the two had a synergistic inhibitory effect on A549/DDP cells.Compared with the blank group,IN group,DDP group,IN+DDP combination group significantly blocked the cell cycle in G0/G1 phase,increased the apoptotic rate of A549/DDP cells(P<0.01),reduced the resistance proteins P-gp and LRP of A549/DDP cells and MRP expression level(P<0.01),and significantly down-regulated the expression level of PI3K and p-AKt protein(P<0.01).Conclusions The combination of IN and DDP can synergistically inhibit the proliferation of A549/DDP cells,arrest the cell cycle and promote cell apoptosis.The mechanism may be related to the inhibition of the expression of drug-resistant proteins P-gp,LRP and MRP,and inhibition of the activity of PI3K/AKt signaling pathway,thereby enhancing the effect of DDP on drug-resistant cells.
作者
郭亚楠
郭红云
王涛
张永东
郭文静
蒋兵
苏海翔
GUO Ya-nan;GUO Hong-yun;WANG Tao;ZHANG Yong-dong;GUO Wen-jing;JIANG Bing;SU Hai-xiang(Gansu Academy Institute for Medical Research, Gansu Cancer Hospital, Lanzhou 730050, China;School of Basic Medicine, Gansu University of Traditional Chinese Medicine, Lanzhou 730000, China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2021年第10期1429-1434,共6页
Chinese Pharmacological Bulletin
基金
甘肃省中医药管理局科研课题(No GKZ-2015-9)。
关键词
黄腐酚
顺铂
肺癌
凋亡
周期
协同作用
xanthohumol
cisplatin
lung cancer
apoptosis
cycle
synergistic effect
