背景:肺癌是癌症死亡的主要原因,非小细胞肺癌(NSCLC)占肺癌相关死亡率的大部分.近年来,已经有许多治疗非小细胞肺癌的方法,但是治愈率和存活率仍然非常低。异巴瓦查尔酮(IBC)属于中药补骨脂的查尔酮成分,是一种独特的蛋白激酶B(AKT)通路抑制剂,具有显著的抗癌作用。以前的研究表明,IBC具有多种生物学特性,包括抗癌,抗炎,和抗氧化性能。本研究主要运用网络药理学分析方法,分子对接技术和实验验证阐明IBC治疗NSCLC的潜在机制。
方法:使用网络药理学筛选NSCLC中IBC作用的关键基因和途径。IBC靶基因来自中医百科全书(ETCM)和BATMAN-TCM数据库,NSCLC靶基因来自GeneCards,在线孟德尔人遗传(OMIM)和治疗目标数据库(TTD)数据库,将两者作为蛋白质-蛋白质相互作用网络分析和富集分析的交叉基因,通过分子对接进一步验证了化合物与核心靶标的结合能。然后进行体外细胞系实验以进一步阐明IBC用于NSCLC的机制。
结果:通过搜索IBC和NSCLC靶标的交集,共检索到279个潜在靶标。蛋白质-蛋白质相互作用(PPI)网络分析表明,6个靶标,包括AKT1,RXRA,NCOA1,RXRB,RARA,PPARG是hub基因。基因本体论(GO)和京都基因和基因组百科全书(KEGG)分析表明IBC治疗NSCLC主要涉及类固醇结合,转录因子活性,癌症的途径,cAMP信号通路,腺苷5'-单磷酸(AMP)激活的蛋白激酶(AMPK)信号通路。其中,AMPK信号通路,其中含有最多的富集基因,可能在NSCLC的治疗中发挥更大的作用。然后,体外实验结果表明,IBC可抑制NSCLC细胞的增殖,诱导细胞自噬和凋亡。结果还表明,IBC可以增加AMPK的蛋白表达,降低AKT和哺乳动物雷帕霉素靶蛋白(mTOR)的表达,提示IBC可以通过诱导细胞自噬和凋亡以及调节AMPK和AKT信号通路来治疗NSCLC。
结论:总之,本研究通过网络药理学和实验验证,为IBC抗NSCLC的保护机制提供了新的思路.
BACKGROUND: Lung cancer is the leading cause of cancer deaths, and non-small cell lung cancer (NSCLC) accounts for the majority of lung cancer-related mortality. In recent years, there have been numerous treatments for non-small cell lung cancer, but the cure and survival rates are still extremely low.
Isobavachalcone (IBC) belongs to the chalcone component of the traditional Chinese medicine Psoralea corylifolia L., and is a unique Protein kinase B (AKT) pathway inhibitor with significant anticancer effects. Previous studies have shown that IBC possess a variety of biological properties, including anti-cancer, anti-inflammatory, and antioxidant properties. This study focused on the use of network pharmacology analysis, molecular docking technology and experimental validation to elucidate the potential mechanisms of IBC for the treatment of NSCLC.
METHODS: Screening key genes and pathways of IBC action in NSCLC using network pharmacology. The IBC target genes were from The Encyclopedia of Traditional Chinese Medicine (ETCM) and BATMAN-TCM databases, the NSCLC target genes were from GeneCards, Online Mendelian Inheritance in Man (OMIM) and The Therapeutic Target database (TTD) databases, both of which were taken as intersecting genes for protein-protein interaction network analysis and enrichment analysis, and the binding energies of the compounds to the core targets were further verified by molecular docking. Cell lines in vitro experiments were then performed to further unravel the mechanism of IBC for NSCLC.
RESULTS: A total of 279 potential targets were retrieved by searching the intersection of IBC and NSCLC targets. Protein-protein interaction (PPI) network analysis indicated that 6 targets, including AKT1, RXRA, NCOA1, RXRB, RARA, PPARG were hub genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis suggested that IBC treatment of NSCLC mainly involves steroid binding, transcription factor activity, Pathways in cancer, cAMP signaling pathway, Adenosine 5\'-monophosphate (AMP)-activated protein kinase (AMPK) signaling pathway. Among them, the AMPK signaling pathway, which contained the largest number of enriched genes, may play a greater role in the treatment of NSCLC. Then, the results of in vitro experiment indicated that IBC could inhibit proliferation of NSCLC cells and induce cell autophagy and apoptosis. The results also showed that IBC could increase the protein expression of AMPK and decrease the protein expression of AKT and mammalian target of rapamycin (mTOR), suggesting that IBC can treat NSCLC by inducing cellular autophagy and apoptosis as well as modulating AMPK and AKT signaling pathways.
CONCLUSIONS: In summary, this study provided a new insight into the protective mechanism of IBC against NSCLC through network pharmacology and experimental validation.