补中益气汤中的有效成分槲皮素通过GSTP1-JAK-STAT途径增强肺腺癌耐药细胞株A549/DDP对顺铂的敏感性  

Study on the Quercetin,An Active Component from the Buzhong Yiqi Decoction Enhances Sensitivity of Lung Adenocarcinoma Resistant Cell Line A549/DDP to Cisplatin via the GSTP1-JAK-STAT Pathway

作　　者：邹长鹏[1] 郭燕 张长旺 高思铭 李娜[1] ZOU Chang-Peng;GUO Yan;ZHANG Chang-Wang;GAO Si-Ming;LI Na(Dept.of Oncology,The First Hospital of Hebei Medical University,Shijiazhuang 050031 Hebei,China)

机构地区：[1]河北医科大学第一医院肿瘤科,河北石家庄050031

出　　处：《广州中医药大学学报》2023年第10期2606-2614,共9页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：河北省中医药管理局课题(编号:2022388)。

摘　　要：【目的】探究补中益气汤的有效成分槲皮素联合顺铂治疗在增强肺腺癌对顺铂敏感性中的作用及其潜在分子机理。【方法】(1)网络药理学分析:从TCMSP数据库筛选补中益气汤的有效成分及其对应靶点,从TCGA数据库下载肺腺癌疾病的数据,并基于差异分析和网络药理学分析挖掘补中益气汤治疗肺腺癌的主要活性成分和潜在作用靶点,通过富集分析以判断相关信号通路,通过分子对接模拟技术分析关键基因与有效成分之间的对接关系。(2)细胞实验:槲皮素或顺铂单一处理以及两者联合处理肺腺癌细胞(A549、A549/DDP),通过细胞计数试剂盒(CCK)-8、Western Blot法、流式细胞术分别对应分析肺腺癌细胞的细胞活力、关键蛋白的表达和细胞凋亡情况。细胞热迁移分析(CETSA)验证活性成分槲皮素与靶蛋白谷胱甘肽S-转移酶π(GSTP1)的结合关系。【结果】补中益气汤中的有效成分槲皮素能与GSTP1紧密结合。通路富集分析显示槲皮素和顺铂可能通过铂类耐药以及Janus激酶(JAK)-信号传导与转录激活因子(STAT)通路影响肺腺癌的进展。此外,槲皮素联合顺铂可下调肺腺癌细胞中GSTP1和JAK-STAT信号通路相关蛋白的表达。在功能上,槲皮素能加强顺铂诱导的肺腺癌细胞的抗癌活性。【结论】槲皮素可减轻肺腺癌耐药细胞对顺铂的耐药性。Objective To investigate the role of quercetin,the active component of Buzhong Yiqi Decoction,in combination with cisplatin treatment in enhancing the sensitivity of lung adenocarcinoma to cisplatin and its potential molecular mechanism.Methods(1)Network pharmacology analysis:the active components of Buzhong Yiqi Decoction and their corresponding targets were screened from TCMSP,data of lung adenocarcinoma patients were downloaded from TCGA database,and the main active components and potential targets of Buzhong Yiqi Decoction for the treatment of lung adenocarcinoma were mined based on difference analysis and network pharmacology analysis,and enrichment analysis was performed to determine the relevant signaling pathways,followed by molecular docking simulation to analyze the docking relationship between key genes and active components.(2)Cell-based assays:lung adenocarcinoma cells(A549,A549/DDP)were treated with quercetin or cisplatin alone or in combination,and cell viability,expression of key proteins and apoptosis of lung adenocarcinoma cells were analyzed by cell counting kit(CCK)-8,Western Blot and flow cytometry,respectively.Cellular thermal shift assay(CETSA)was performed to verify the interaction of active component quercetin on target protein glutathione S-transferaseπ(GSTP1).Results Quercetin,the active component in cisplatin,binds tightly to GSTP1.Pathway enrichment analysis revealed that quercetin and cisplatin may affect lung adenocarcinoma progression through platinum resistance as well as the Janus kinase(JAK)-signaling and signal transducers and activators of transcription(STAT)pathway.In addition,quercetin combined with cisplatin down-regulated the expressions of GSTP1 and JAK-STAT signaling pathway-related proteins in lung adenocarcinoma cells.Functionally,quercetin enhanced the anticancer activity of cisplatin-induced lung adenocarcinoma cells.Conclusion Quercetin attenuated the drug resistance to cisplatin in lung adenocarcinoma resistant cells.

关 键 词：补中益气汤 槲皮素 肺腺癌 GSTP1 JAK-STAT途径 顺铂敏感性 A549/DDP细胞 

分 类 号：R285.5[医药卫生—中药学]