Computational and In Vitro Analysis of Plumbagin’s Molecular Mechanism for the Treatment of Hepatocellular Carcinoma

Abstract

Hepatocellular carcinoma (HCC) is the fifth most common malignant tumor and the second leading cause of cancer-related death in the world. Plumbagin (PL) is a small molecule naphthoquinone compound isolated from Plumbago zeylanica L. that has important anticancer properties, but its mechanism requires further investigation. In this study, we used a comprehensive network pharmacology approach to study the mechanism of action of PL for the treatment of HCC. The method includes the construction of multiple networks; moreover, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to identify biological processes and signaling pathways. Subsequently, in vitro experiments were performed to verify the predicted molecular mechanisms obtained from the network pharmacology-based analysis. Network pharmacological analysis showed that PL may exert anti-HCC effects by enhancing reactive oxygen species (ROS) production to generate oxidative stress and by regulating the PI3K/Akt and MAPK signaling pathways. In vitro experiments confirmed that PL mainly mediates the production of ROS, regulates the PI3K/Akt and MAPK signaling pathways to promote apoptosis and autophagy, and shows significant therapeutic effects on HCC. In conclusion, our work proposes a comprehensive systems pharmacology approach to explore the potential mechanism of PL for the treatment of HCC.

Plumbagin Exhibits Genotoxicity and Induces G2/M Cell Cycle Arrest via ROS-Mediated Oxidative Stress and Activation of ATM-p53 Signaling Pathway in Hepatocellular Cells

“…It was previously demonstrated that plumbagin inhibits the proliferation of HCC cell lines such as SMMC-7721, BEL-7404 and LM3 [ 16 , 24 ]. To select the working concentrations, Huh-7 and Hep-G2 cells were treated by PLB at different concentrations for 12 h. Cell counting kit-8 (CCK-8) analysis results revealed that the concentration dependently inhibited the proliferation of HCC cells with IC 50 values of 11.49 μM for Huh-7 cells and 16.42 μM for Hep-G2 cells ( Figure 1 A,B).…”

“…Furthermore, the level of cellular antioxidants such as glutathione rapidly decreased after PLB treatment, indicating excessive oxidative stress in HCC cells ( Figure 3 B–E). Recently, a computational analysis has shown that plumbagin may exert anti-HCC effects by enhancing ROS production and function as an upstream signal factor that regulates the PI3K/Akt/mTOR pathway, which is of critical importance in plumbagin-induced cell apoptosis and autophagy [ 16 ]. In lung cancer A549 cells, plumbagin could engender apoptosis via caspase-9 activation and ROS production [ 39 ].…”

“…ROS have been reported to play an important role in PLB-induced cytotoxicity, apoptosis and autophagy [ 16 , 23 ]. NAC was also found to protect against growth inhibition using plumbagin [ 46 ], indicating that ROS play a key role in PLB’s anticancer effects.…”

“…In HCC, previous studies have shown that plumbagin could induce autophagy and apoptosis in HCC cells in vitro and in vivo, and its proapoptotic ability was found to be related to caspase 3/vimentin-mediated epithelial–mesenchymal transition (EMT) [ 14 , 15 ]. Recently, a network-pharmacology-based study delineated the importance of reactive oxygen species (ROS) production and the accompanying oxidative stress regarding PLB’s anti-HCC effects [ 16 ]. Nevertheless, a comprehensive anti-HCC pharmacological mechanism, especially the fundamental role played by the ROS-induced oxidative stress triggered by PLB, remains unclear.…”

Systems Pharmacology-Based Strategy to Investigate the Mechanism of Ruangan Lidan Decoction for Treatment of Hepatocellular Carcinoma

“…All gene names were entered into the DAVID database and limited the species to Homo sapiens . Only those terms with the P value < 0.05 were retained [ 29 ].…”