Tan Xi-Ying, Tao Jing, Zhang Yu, Gu Ru-Xin
Department of Pharmacy, Affiliated Hospital of Nanjing University of Chinese Medicine Nanjing 210029, China.
China Pharmaceutical University Nanjing 210009, China.
Zhongguo Zhong Yao Za Zhi. 2023 Sep;48(18):5056-5067. doi: 10.19540/j.cnki.cjcmm.20230601.701.
This study aims to investigate the mechanism of Astragali Radix-Curcumae Rhizoma(HQEZ) in the treatment of gastric cancer based on network pharmacology. Further, the SGC7901 cell model of gastric cancer was employed to validate the efficacy and key targets of the herb pair. Firstly, the CCK-8 assay was employed to evaluate the direct effect of HQEZ on the proliferation of gastric cancer SGC7901 cells. Then, network pharmacology methods were employed to investigate the active ingredients, key targets, and key signaling pathways involved in the treatment of gastric cancer with HQEZ. The results showed that HQEZ contained 18 potential active ingredients, such as quercetin, naringenin, and curcumin. The results of gene ontology(GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment suggested that the main targets of HQEZ in treating gastric cancer were involved in the regulation of protein serine/threonine kinase activity, activation of mitogen-activated protein kinase(MAPK) activity, cysteine-type endopeptidase activity, and negative regulation of protein serine/threonine kinase activity. The hypoxia-inducible factor-1(HIF-1) signaling pathway, ATP-binding cassette(ABC) transporters, cytochrome P450-mediated metabolism of xenobiotics, p53 signaling pathway, and cell apoptosis were key signaling pathways of HQEZ in treating gastric cancer. The cell experiments demonstrated that HQEZ significantly downregulated the expression of ATP-binding cassette subfamily B member 1(ABCB1), epidermal growth factor receptor(EGFR), phosphorylated serine/threonine kinase(p-AKT), hypoxia inducible factor 1 subunit alpha(HIF1A), B-cell lymphoma 2(BCL2), breast cancer susceptibility protein 1(BRCA1), DNA polymerase theta(POLH), ribonucleotide reductase M1(RRM1), and excision repair cross-complementation group 1(ERCC1), and upregulated the expression of tumor protein P53(TP53) and cysteinyl aspartate-specific proteinase(CAPS3). Finally, a multivariate COX regression model was adopted to study the relationship between gene expression and clinical information data of gastric cancer patients in the TCGA database, which demonstrated that the key targets of HQEZ were associated with the poor prognosis in gastric cancer patients. Further feature selection using the LASSO algorithm showed that EGFR, HIF1A, TP53, POLH, RRM1, and ERCC1 were closely associated with the survival of gastric can-cer patients. In conclusion, HQEZ regulates the expression of genes involved in DNA repair, survival, and apoptosis in gastric cancer cells via multiple targets and pathways, assisting the treatment of gastric cancer.
本研究旨在基于网络药理学探讨黄芪-莪术药对（HQEZ）治疗胃癌的作用机制。此外，采用胃癌SGC7901细胞模型验证该药对的疗效及关键靶点。首先，采用CCK-8法评估HQEZ对胃癌SGC7901细胞增殖的直接影响。然后，运用网络药理学方法研究HQEZ治疗胃癌所涉及的活性成分、关键靶点及关键信号通路。结果显示，HQEZ含有槲皮素、柚皮素、姜黄素等18种潜在活性成分。基因本体（GO）功能注释和京都基因与基因组百科全书（KEGG）通路富集结果表明，HQEZ治疗胃癌的主要靶点涉及蛋白丝氨酸/苏氨酸激酶活性的调节、丝裂原活化蛋白激酶（MAPK）活性的激活、半胱氨酸型内肽酶活性以及蛋白丝氨酸/苏氨酸激酶活性的负调节。缺氧诱导因子-1（HIF-1）信号通路、ATP结合盒（ABC）转运体、细胞色素P450介导的外源性物质代谢、p53信号通路和细胞凋亡是HQEZ治疗胃癌的关键信号通路。细胞实验表明，HQEZ显著下调ATP结合盒亚家族B成员1（ABCB1）、表皮生长因子受体（EGFR）、磷酸化丝氨酸/苏氨酸激酶（p-AKT）、缺氧诱导因子1亚基α（HIF1A）、B细胞淋巴瘤2（BCL2）、乳腺癌易感蛋白1（BRCA1）、DNA聚合酶θ（POLH）、核糖核苷酸还原酶M1（RRM1）和切除修复交叉互补组1（ERCC1）的表达，并上调肿瘤蛋白P53（TP53）和半胱天冬酶（CAPS3）的表达。最后，采用多变量COX回归模型研究TCGA数据库中胃癌患者基因表达与临床信息数据之间的关系，结果表明HQEZ的关键靶点与胃癌患者的不良预后相关。使用LASSO算法进一步进行特征选择显示，EGFR、HIF1A、TP53、POLH、RRM1和ERCC1与胃癌患者的生存密切相关。综上所述，HQEZ通过多个靶点和通路调节胃癌细胞中参与DNA修复、生存和凋亡的基因表达，辅助治疗胃癌。
