Liu Jing, Yang Yan, Zeng Yan, Qin Xiang, Guo Ling, Liu Wenjun
Department of Pediatrics, Children Hematological Oncology and Birth Defects Laboratory, The Affiliated Hospital of Southwest Medical University, Sichuan Clinical Research Center for Birth Defects, Luzhou, Sichuan, 646000, China.
Heliyon. 2023 Feb 24;9(3):e14009. doi: 10.1016/j.heliyon.2023.e14009. eCollection 2023 Mar.
To explore the mechanism of PG against acute lymphoblastic leukaemia (ALL) by network pharmacology and experimental verification in vitro.
First, the biological activity of PG against B-ALL was determined by CCK-8 and flow cytometry. Then, the potential targets of PG were obtained from the PharmMapper database. ALL-related genes were collected from the GeneCards, OMIM and PharmGkb databases. The two datasets were intersected to obtain the target genes of PG in ALL. Then, protein interaction networks were constructed using the STRING database. The key targets were obtained by topological analysis of the network with Cytoscape 3.8.0 software. In addition, the mechanism of PG in ALL was confirmed by protein‒protein interaction, gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses. Furthermore, molecular docking was carried out by AutoDock Vina. Finally, Western blotting was performed to confirm the effect of PG on NALM6 cells.
PG inhibited the proliferation of NALM6 cells. A total of 174 antileukaemic targets of PG were obtained by network pharmacology. The key targets included AKT1, MAPK14, EGFR, ESR1, LCK, PTPN11, RHOA, IGF1, MDM2, HSP90AA1, HRAS, SRC and JAK2. Enrichment analysis found that PG had antileukaemic effects by regulating key targets such as MAPK signalling, and PG had good binding activity with MAPK14 protein (-8.9 kcal/mol). PG could upregulate the expression of the target protein p-P38, induce cell cycle arrest, and promote the apoptosis of leukaemia cells.
MAPK14 was confirmed to be one of the key targets and pathways of PG by network pharmacology and molecular experiments.
通过网络药理学和体外实验验证，探讨PG抗急性淋巴细胞白血病（ALL）的机制。
首先，采用CCK-8法和流式细胞术检测PG对B-ALL的生物学活性。然后，从PharmMapper数据库中获取PG的潜在靶点。从GeneCards、OMIM和PharmGkb数据库中收集ALL相关基因。将这两个数据集进行交集分析，以获得PG在ALL中的靶基因。然后，使用STRING数据库构建蛋白质相互作用网络。通过Cytoscape 3.8.0软件对网络进行拓扑分析，获得关键靶点。此外，通过蛋白质-蛋白质相互作用、基因本体论和京都基因与基因组百科全书通路富集分析，证实PG在ALL中的作用机制。进一步，使用AutoDock Vina进行分子对接。最后，进行蛋白质印迹法以确认PG对NALM6细胞的作用。
PG抑制NALM6细胞的增殖。通过网络药理学共获得174个PG的抗白血病靶点。关键靶点包括AKT1、MAPK14、EGFR、ESR1、LCK、PTPN11、RHOA、IGF1、MDM2、HSP90AA1、HRAS、SRC和JAK2。富集分析发现，PG通过调节MAPK信号等关键靶点发挥抗白血病作用，且PG与MAPK14蛋白具有良好的结合活性（-8.9 kcal/mol）。PG可上调靶蛋白p-P38的表达，诱导细胞周期阻滞，并促进白血病细胞凋亡。
通过网络药理学和分子实验证实MAPK14是PG的关键靶点和通路之一。
