摘要
目的:探究补中益气汤联合顺铂通过调控Wnt/β-catenin信号通路改善A549/DDP细胞耐药性的分子机制。方法:培养A549/DDP细胞并制备补中益气汤含药血清。采用siRNA沉默GSK-3β表达,检测各组A549/DDP细胞中GSK-3β蛋白的相对表达量;Western blot检测沉默GSK-3β后各组A549/DDP细胞β-catenin和Survivin表达;MTT法检测GSK-3β受抑对各组A549/DDP细胞顺铂IC_(50)的影响;采用Annexin V/PI染色法检测GSK-3β受抑对各组A549/DDP细胞凋亡的影响。结果:相较于空白对照组,siGSK-3β-592组对蛋白表达的抑制率达70%(P<0.05);RNAi技术沉默GSK-3β后,A549/DDP细胞中β-catenin和Survivin蛋白表达明显增加(P<0.05),单独顺铂治疗组与单独补中益气汤治疗组均可使β-catenin和Survivin蛋白表达明显下调(P<0.05),补中益气汤联合顺铂治疗组则可使β-catenin和Survivin蛋白表达下调更显著(P<0.05);RNAi技术沉默GSK-3β后,A549/DDP细胞顺铂IC_(50)显著升高(P<0.05),而补中益气汤仍可显著降低A549/DDP细胞的顺铂IC_(50)(P<0.05);GSK-3β表达受抑后,A549/DDP细胞的顺铂诱导凋亡率明显受抑制(P<0.05),而补中益气汤仍可明显提高A549/DDP细胞的顺铂诱导凋亡率(P<0.05)。结论:上调A549/DDP细胞中Wnt/β-catenin信号通路活性时,补中益气汤联合顺铂仍对A549/DDP细胞的顺铂耐药性有明显改善作用。
Objective:To investigate molecular mechanism of Buzhong Yiqi Decoction combined with cisplatin in improving drug resistance of A549/DDP cells by regulating Wnt/β-catenin signaling pathway.Methods:A549/DDP cells were cultured,and the containing Buzhong Yiqi Decoction was prepared.siRNA was used to silence expression of GSK-3β,and relative expression of GSK-3βin A549/DDP cells was detected.Expressions ofβ-catenin and Survivin in A549/DDP cells of each group after GSK-3βsilenced were detected by Western blot.MTT assay was used to detect effect of GSK-3βinhibition on cisplatin IC_(50) in A549/DDP cells of each group.Annexin V/PI staining was used to detect effect of GSK-3βinhibition on apoptosis of A549/DDP cells in each group.Results:Compared with blank control group,inhibitory rate of siGSK-3β-592 group on protein expression was 70%(P˂0.05);after silencing GSK-3βby RNAi technology,expressions ofβ-catenin and Survivin in A549/DDP cells were increased significantly(P˂0.05),protein expressions ofβ-catenin and Survivin were significantly decreased in both cisplatin and Buzhong Yiqi Decoction groups(P˂0.05),and the combination of Buzhong Yiqi Decoction and cisplatin could even significantly decrease expressions ofβ-catenin and Survivin(P˂0.05);after silencing GSK-3βby RNAi technology,cisplatin IC_(50) of A549/DDP cells was increased significantly(P˂0.05),while Buzhong Yiqi Decoction could still significantly reduce cisplatin IC_(50) of A549/DDP cells(P˂0.05);after expression of GSK-3βwas inhibited,apoptosis rate of A549/DDP cells induced by cisplatin was inhibited obviously(P˂0.05),while apoptosis rate of A549/DDP cells induced by cisplatin was still increased significantly by Buzhong Yiqi Decoction(P˂0.05).Conclusion:Cisplatin resistance of A549/DDP cells can still be improved by Buzhong Yiqi Decoction combined with cisplatin when activity of Wnt/β-catenin signaling pathway was up-regulated in A549/DDP cells.
作者
牟琪瑞
刘春英
王哲
王莹
于丹
潘茜
井欢
高原(指导)
MU Qirui;LIU Chunying;WANG Zhe;WANG Ying;YU Dan;PAN Qian;JING Huan;GAO Yuan(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2023年第5期961-965,972,共6页
Chinese Journal of Immunology
基金
国家自然科学基金项目(81403303,81973735)
辽宁省教育厅科学研究经费项目(面上项目)(LJKZ0901)。