芦丁对人肺癌A549/DDP细胞耐药性的逆转作用及其机制  

Reversal effect of rutin on cisplatin-resistance in human lung cancer A549/DDP cells and its mechanisms

作　　者：孔祥虎[1] 李志欣 房丽君[1] 焦建峰[1] KONG Xiang-Hu;LI Zhi-Xin;FANG Li-Jun;JIAO Jian-Feng(Baotou Tumor Hospital,Baotou 014030,China)

机构地区：[1]包头市肿瘤医院,包头014030 [2]包钢医院,包头014010

出　　处：《中国免疫学杂志》2019年第19期2332-2336,共5页Chinese Journal of Immunology

摘　　要：目的:本研究旨在观察芦丁(RT)对人肺癌A549/DDP细胞顺铂(DDP)耐药性的影响,并探讨其可能的分子机制。方法:体外常规培养A549细胞和A549/DDP细胞,给予不同剂量RT和/或DDP处理48 h后,采用CCK-8法、Chou-Talalay中效分析法、流式细胞术、免疫印迹法和实时定量PCR反应分别检测了细胞活力、药物联合效应、细胞凋亡、IKKα和p65蛋白磷酸化水平以及MRP1和GST-π蛋白和mRNA表达水平。结果:DDP可剂量依赖性地抑制A549细胞和A549/DDP细胞活力(P<0.05),A549/DDP细胞耐药倍数为7.49;RT可协同增强DDP对A549/DDP细胞的抑制作用,逆转倍数为2.33;与对照组相比,DDP组A549/DDP细胞凋亡率显著升高(P<0.05),而低毒剂量的RT可协同增强DDP引起的A549/DDP细胞凋亡(P<0.05);同时,RT和IKK16均可抑制A549/DDP细胞中IKKα和p65蛋白磷酸化(P<0.05),下调MRP1和GST-π的mRNA和蛋白表达水平(P<0.05),且两者联用时抑制作用增强(P<0.05)。结论:RT可通过抑制IKKα/p65通路,进而下调MRP1和GST-π表达,以逆转A549/DDP细胞顺铂耐药性。Objective:To investigate the effect of rutin(RT)on cisplatin(DDP)resistance in human lung cancer A549/DDP cells and its possible mechanism.Methods:A549 and A549/DDP cells were cultured in vitro and treated with RT and/or DDP at different concentrations for 48 h and then the cell viability were detected by CCK-8 assay.The combination index(CI)of RT and DDP were analyzed by Chou-Talalay method.The cell apoptosis were observed by flow cytometry.Western blot was performed to analyze the protein levels of IKKα,p-IKKα(Ser176),p65,p-p65(Ser536),MRP1 and GST-π.The mRNA levels of MRP1 and GST-πwere detected by RT-PCR.Results:DDP remarkably inhibited the cell activity of A549 and A549/DDP cells in dose-dependent manner,and the resistant factor of A549/DDP cells was 7.49.Co-treatment with RT and DDP produced a synergistic effect with a reversal fold of 2.33.DDP remarkably induced apoptosis in A549/DDP cells(P<0.05).The effect was enhanced by RT under a non-toxic concentration(P<0.05).Meanwhile,RT or IKK16 significantly inhibited the phosphorylation of IKKαand p65,and down-regulated the protein and mRNA levels of MRP1 and GST-πin A549/DDP cells(P<0.05).The co-treatment with RT and DDP induced a lower expression of MRP1 and GST-πthan RT alone(P<0.05).Conclusion:RT may enhance the sensibility of A549/DDP cells to DDP by suppressing the phosphorylation of IKKαand p65,and then down-regulated the protein and mRNA levels of MRP1 and GST-π.

关 键 词：芦丁 A549/DDP细胞 IκB激酶α P65 耐药 

分 类 号：R734.2[医药卫生—肿瘤]