Integrating Network Pharmacology and Bioinformatics to Explore the Effects of Dangshen (Codonopsis pilosula) Against Hepatocellular Carcinoma: Validation Based on the Active Compound Luteolin

Yaping Yu,1,* Shun Ding,2,* Xiaoqing Xu,1,* Dongming Yan,3 Yonghao Fan,1 Banzhan Ruan,1 Xiaodian Zhang,1 Liping Zheng,1 Wei Jie,1 Shaojiang Zheng1,4 1Department of Oncology of the First Affiliated Hospital & Tumor Institute, Hainan Medical University, Haikou, 570102, People’s Republic of China; 2Department of Otolaryngology, Head and Neck Surgery, the First Affiliated Hospital, Hainan Medical University, Haikou, 570102, People’s Republic of China; 3Department of Neurosurgery, the First Affiliated Hospital of Hainan Medical University, Haikou, 570102, People’s Republic of China; 4Key Laboratory of Emergency and Trauma, Ministry of Education, Hainan Medical University, Haikou, 571199, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wei Jie; Shaojiang Zheng, Department of Oncology of the First Affiliated Hospital & Tumor Institute, Hainan Medical University, Haikou, 570102, People’s Republic of China, Tel +86-898-66968217 ; +86-898-66568158, Email wei_jie@hainmc.edu.cn; zhengsj2008@163.comPurpose: This study aimed to explore the pharmacological mechanism of Dangshen (Codonopsis pilosula) against hepatocellular carcinoma (HCC) based on network pharmacology and bioinformatics, and to verify the anticancer effect of luteolin, the active ingredient of Codonopsis pilosula, on HCC cells.Methods: The effective compounds and potential targets of Codonopsis pilosula were established using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) database. The genes related to HCC were obtained through the GeneCards database. The interactive genes were imported into the Visualization and Integrated Discovery database for Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) signal enrichment, and the hub genes were screened out. The Cancer Genome Atlas database was used to construct a prognosis model, and the prognosis and clinicopathological correlation were analyzed. In in vitro experiments, we verified the effects of luteolin, an active compound of Codonopsis pilosula, on the proliferation, cell cycle, apoptosis and migration of HCC cells.Results: A total of 21 effective compounds of Codonopsis pilosula and 98 potential downstream target genes were screened through the TCMSP database, and 1406 HCC target genes were obtained through the GeneCards database. Finally, 53 interacting genes between the two databases were obtained, among which, the 10 key node genes were CASP3, TP53, MDM2, AKT1, ESR1, BCL2L1, MCL1, HSP90AA1, CASP9, and CCND1, involving 77 typical GO terms and 72 KEGG signals. The Kaplan–Meier survival curve of the model group showed that the overall survival of the low-risk group was significantly higher than that of the high-risk group. Luteolin significantly inhibited the proliferation and migration of HCC cells, induced apoptosis, and increased the G2/M phase ratio. Mechanistically, luteolin significantly inhibited the phosphorylation of MAPK-JNK and Akt (Thr308) and subsequently led to upregulation of ESR1. Pharmacological inhibition of ESR1 with fulvestrant enhanced cell viability and migration and attenuated apoptosis.Conclusion: Codonopsis pilosula has potential for clinical development due to its anti-HCC properties. Luteolin, the effective component of Codonopsis pilosula, plays anti-HCC role through AKT- or MAPK-JNK signaling mediated ESR1.Keywords: Codonopsis pilosula, network pharmacology, bioinformatics, luteolin, hepatocellular carcinoma