药用植物酒精浸提组方AEAMP9对肺癌A549细胞的抑制作用及机制

Inhibition and mechanism of AEAMP9,a novel compound formula of alcoholic extract of anticancer medicinal plants,on lung cancer A549 cells

研究以体外培养的A549肺癌细胞为靶标,通过对124种可能与抗癌活性相关的植物源中药材酒精提取物的抗肿瘤作用规模化筛选,获得各中药材抗A549细胞活性的排序,研发了一种安全有效和新的抗肿瘤药用植物组方AEAMP9(获国家知识产权局发明专利授权)并探索其可能的分子作用机制。AEAMP9由白花蛇舌草、黄柏、甘草、侧柏、野菊花和羌活等6种常用中药材的酒精提取物配伍制备而成。研究发现,AEAMP9可以有效抑制A549的细胞活力、克隆形成与细胞迁移,并促进细胞凋亡。AEAMP9显著降低ABCG2、FoxM1、Vimentin、MMP14基因的表达和Bcl-2/Bax比值,促进凋亡相关基因p53、caspase 3、caspase 8、caspase 9的表达。网络药理学和细胞RNA转录组测序结果揭示:TNF、IL-17等与肿瘤、炎症、凋亡相关的重要信号通路受到调控,EGR1、IL-6、ALK、VEGF可能是AEAMP9-肺腺癌作用网络的关键调控靶点,AEAMP9中的活性成分槲皮素、山柰酚、木犀草素可能在其抗肿瘤活性中发挥重要的作用。AEAMP9对昆明小鼠无明显的急性毒性作用,其对A549细胞移植瘤小鼠模型肿瘤生长具有显著的抑制作用。

In view of the high morbidity and mortality of lung cancer,lung cancer was concerned and human non-small cell lung cancer A549 cells were used as the target in this study.Here,after screening 124 kinds of medicinal plants in A549 lung cancer cells and the determination of their anticancer orders,an efficient and novel compound formula of AEAMP9 was developed and its underlying molecular mechanism was investigated.AEAMP9 was prepared from the alcoholic extracts of six anticancer medicinal plants including Hedyotis diffusa Willd.(Bai Hua She She Cao),Phellodendron amurense Rupr.(Huang Bai),Glycyrrhiza uralensis Fisch.(Gan Cao),Platycladus orientalis(L.)Franco(Ce Bai Ye),Chrysanthemum indicum L.(Ye Ju Hua)and Notopterygium incisum Ting ex H.T.Chang(Qiang Huo).It can effectively inhibit the viability and clone formation of cancer cells,provoke apoptosis and block migration in A549 cells.Mechanistically,AEAMP9 significantly decreased the expressions of ABCG2,FoxM1,Vimentin,MMP14 and the ratio of Bcl-2/Bax,consequently promoted the activation of apoptosis associated genes including p53,caspase 3,caspase 8 and caspase 9.Network pharmacology and transcriptome RNA-seq revealed that many genes were significantly and differentially expressed in A549 cells after treatment with AEAMP9.TNF,IL-17 and other important signal pathways related to tumor,inflammation and apoptosis were closely regulated.EGR1,IL-6,ALK and VEGF may be the key regulatory targets for the interaction of AEAMP9 and lung adenocarcinoma.The components of quercetin,kaempferol and luteolin in AEAMP9 may play an important role in antitumor activity.Safely,AEAMP9 had no obvious toxic effects on SPF Kunming mice.Furthermore,AEAMP9 can significantly inhibit the tumor growth in mouse A549 xenograft models.