黄连素和吴茱萸碱协同抗癌的miRNA网络机制研究

摘要

目的 从miRNA 及其调控的通路网络水平,研究黄连素和吴茱萸碱产生协同抗癌作用的机制. 方法 先在细胞水平上验证并详细分析了黄连素与吴茱萸碱对肿瘤细胞增殖的抑制作用,确定了二者的剂量效应范围及协同作用产生的浓度配比,然后采用基因表达芯片技术分析了黄连素和吴茱萸碱处理后的肝癌细胞BEL-7402的miRNA表达谱,通过构建miRNA-mRNA互作网络,对二者的协同作用机制进行阐释. 结果 黄连素与吴茱萸碱合用可以明显协同抑制肝癌细胞BEL-7402的增殖能力;合用后产生的差异miRNA(DEmiRNA) 主要参与MAPK信号通路、内吞通路及胰岛素信号通路等癌症增殖相关通路的调控;合用影响的特异的靶基因既涵盖了通路上游细胞膜上的3类膜受体,又参与到下游信号通路的转导.结论 从miRNA的调控关系可以看出,黄连素可能在两药协同抑癌作用中起着主要的作用. 黄连素和吴茱萸碱在miRNA水平上协同机制的阐释,为今后协同药物组合的发现提供了一种新的思路.%Aim To explore the mechanisms of action (MOA) of synergistic anticancer function in the combination of berberine and evodiamine.Methods We first analyzed the action of suppression in the drug combination from the cell level and validated the dose scope as well as ratio of concentration in synergistic effects of drug combination.Then, the miRNA chip of liver cancer cell BEL-7402 under different treatment was analyzed.By building the miRNA-mRNA network, the MOA of the synergistic drug combination was illustrated.Results Berberine and evodiamine used in combination could significantly synergistically suppress the proliferative ability of liver cancer cells.The special differentially expressed miRNAs (DEmiRNAs) mainly participated in some cancer proliferation-related pathways and biological processes, such as MAPK signaling pathway, endocytosis pathway and insulin signaling pathway.The special target genes influenced by the drug combination not only covered three kinds of membrane receptors, but also took part in the regulation of downstream pathways.Conclusions From the regulation of miRNAs, it is clear that berberine may play a primary role in the synergistical suppression activity of the drug combination in cancer cells.The discovery of synergistic MOA in the combination of berberine and evodiamine from the miRNA level will provide a new guidance to explore more synergistic drug combinations in the future.