基于非标记定量蛋白质组学技术分析消岩汤干预人肺腺癌细胞后蛋白表达谱特征的影响  

Analyzing the effect of protein expression profile characteristics after the intervention of Xiaoyan Decoction in human lung adenocarcinoma cells based on label-free quantitative proteomics technology

作　　者：郑旭[1,2] 韩燕燕 高珊[1,2] 李翀 Zheng Xu;Han Yanyan;Gao Shan;Li Chong(Department of Pathology,First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300381,China;National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300381,China)

机构地区：[1]天津中医药大学第一附属医院病理科,300381 [2]国家中医针灸临床医学研究中心,天津300381

出　　处：《中华针灸电子杂志》2024年第1期18-23,共6页Chinese Journal of Acupuncture and Moxibustion（Electronic Edition）

基　　金：天津市教委科研计划项目(2018KJ038)。

摘　　要：目的探讨扶正祛瘀解毒法的消岩汤作用于人肺腺癌细胞后对蛋白表达谱特征的影响,并采用生物信息学富集分析差异信号通路。方法以消岩汤干预人肺腺癌A549细胞,提取对照组和消岩汤组细胞的总蛋白,利用非标记定量蛋白质组学技术筛选出差异表达蛋白,应用基因本体论(GO)和应用京都基因与基因组百科全书(KEGG)数据库对差异蛋白及信号通路进行富集分析,并采用Western blot对差异蛋白进行验证。结果与对照组比较,消岩汤组鉴别出1217个差异表达蛋白,上调蛋白384个,下调蛋白833个。通过GO分析发现,差异表达蛋白参与了翻译、mRNA剪接、代谢等生物学过程。KEGG数据库分析显示,差异表达蛋白主要参与前20条信号通路,上调最显著的是核糖体通路,下调最显著的是RNA剪接体和MAPK通路。差异表达蛋白参与肿瘤关系密切的为MAPK通路。Western blot结果显示,与对照组比较,消岩汤组PKA、GRB2、ERK蛋白表达明显下调(t=9.913、9.641、20.060,P均<0.05),其结果与蛋白组学结果具有一致性。结论消岩汤抗肺腺癌作用可能是通过抑制RNA剪接相关的信号及MAPK通路来实现的。Objective To explore the effect of Xiaoyan Decoction,a classic prescription of strengtheing the body,removing blood stasis and detoxifying,on the protein expression profile characteristics of human lung adenocarcinoma cells,and to analyze the differentially expressed signaling pathways using bioinformatics enrichment.Methods Human lung adenocarcinoma A549 cells were treated with Xiaoyan Decoction,total proteins were extracted from the cells of control group and Xiaoyan Decoction,and the differentially expressed proteins were screened by label-free quantitative proteomics.The differentially expressed proteins and signaling pathways were enriched by applying Gene Ontology(GO)database and Kyoto Encyclopedia of Genes and Genomes(KEGG)database,and the differentially expressed proteins were verified by Western blot.Results Compared with the control group,among the 1217 differentially expressed proteins,384 up-regulated proteins and 833 down-regulated proteins were identified in Xiaoyan Decoction group.The GO analysis revealed that the differentially expressed proteins were involved in translation,mRNA splicing,metabolism and other biological processes,and the KEGG database analysis showed that the differentially expressed proteins were mainly involved in the top 20 signaling pathways,with the most significant up-regulation of the ribosomal pathway and the most significant down-regulation of the RNA spliceosome and MAPK pathways.The differentially expressed proteins involved in the tumor were closely related to the MAPK pathway.The results of western blot showed that compared with the control group,PKA,GRB2,and ERK protein expression was significantly down-regulated in the Xiaoyan Decoction group(t=9.913,9.641,20.060,all P<0.05),and the results were consistent with the proteomic results.Conclusion The effect of Xiaoyan Decoction on lung adenocarcinoma may be realized by inhibiting RNA splicing related signaling pathway and MAPK pathway.

关 键 词：肺腺癌 计算生物学 蛋白质组学 消岩汤 MAPK通路 

分 类 号：R285[医药卫生—中药学]