Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2019 Oct 9;10:1185.
doi: 10.3389/fphar.2019.01185. eCollection 2019.

Integrating Network Pharmacology and Pharmacological Evaluation for Deciphering the Action Mechanism of Herbal Formula Zuojin Pill in Suppressing Hepatocellular Carcinoma

Affiliations
Free PMC article

Integrating Network Pharmacology and Pharmacological Evaluation for Deciphering the Action Mechanism of Herbal Formula Zuojin Pill in Suppressing Hepatocellular Carcinoma

Wei Guo et al. Front Pharmacol. .
Free PMC article

Abstract

Hepatocellular carcinoma (HCC) is a kind of complicated disease with an increasing incidence all over the world. A classic Chinese medicine formula, Zuojin pill (ZJP), was shown to exert therapeutic effects on HCC. However, its chemical and pharmacological profiles remain to be elucidated. In the current study, network pharmacology approach was applied to characterize the action mechanism of ZJP on HCC. All compounds were obtained from the corresponding databases, and active compounds were selected according to their oral bioavailability and drug-likeness index. The potential proteins of ZJP were obtained from the traditional Chinese medicine systems pharmacology (TCMSP) database and the traditional Chinese medicine integrated database (TCMID), whereas the potential genes of HCC were obtained from OncoDB.HCC and Liverome databases. The potential pathways related to genes were determined by gene ontology (GO) and pathway enrichment analyses. The compound-target and target-pathway networks were constructed. Subsequently, the potential underlying action mechanisms of ZJP on HCC predicted by the network pharmacology analyses were experimentally validated in HCC cellular and orthotopic HCC implantation murine models. A total of 224 components in ZJP were obtained, among which, 42 were chosen as bioactive components. The compound-target network included 32 compounds and 86 targets, whereas the target-pathway network included 70 proteins and 75 pathways. The in vitro and in vivo experiments validated that ZJP exhibited its prominent therapeutic effects on HCC mainly via the regulation of cell proliferation and survival though the EGFR/MAPK, PI3K/NF-κB, and CCND1 signaling pathways. In conclusion, our study suggested combination of network pharmacology prediction with experimental validation may offer a useful tool to characterize the molecular mechanism of traditional Chinese medicine (TCM) ZJP on HCC.

Keywords: Zuojin pill; cell proliferation and survival; hepatocellular carcinoma; network pharmacology; pharmacological evaluation.

Figures

Figure 1
Figure 1
The technical strategy of the current study.
Figure 2
Figure 2
The compound-target network for ZJP on HCC. The purple nodes represent candidate active compounds and the green nodes represent potential protein targets. The edges represent the interactions between them and nodes size are proportional to their degree.
Figure 3
Figure 3
The 15 most significance of gene ontology (A) and pathway enrichment (B) analysis of therapy target genes of ZJP on HCC.
Figure 4
Figure 4
The target-pathway network for ZJP on HCC. The blue nodes represent targets and the red nodes represent pathways. The edges represent the interactions between them and node size is proportional to their degree.
Figure 5
Figure 5
ZJP inhibited HCC cell growth in Vitro. (A) Time- and dose-dependent effects of ZJP treatment on the viability of HCC cells. (B) Representatives images of colony formation of MHCC97L and PLC/PRF/5 cells. (C) The representative images and statistical graphs of MHCC97L and PLC/PRF/5 cell cycle analysis. (D) The representative images and statistical graphs of migration assay of MHCC97L and PLC/PRF/5 cells. (E) The representative images and statistical graphs of transwell chambers of MHCC97L and PLC/PRF/5 cells. *P < 0.05, **P < 0.01, ***P < 0.001 versus the nontreated group.
Figure 6
Figure 6
ZJP-inhibited tumor growth of orthotopic HCC implantation murine model in vivo. (A) The representative images and statistical graph of luciferase signal of animals throughout the oral treatment. (B) The representative images and statistical graph of tumor size at the end of experiment. (C) The body weight of animals throughout the experiment. (D) ZJP suppressed the invasion of the orthotopic tumor cells into the livers. (E) ZJP suppressed the mitotic events in tumors. *P < 0.05, **P < 0.01, ***P < 0.001 versus the nontreated group.
Figure 7
Figure 7
The relative expressions of related proteins with ZJP treatment on HCC cells (A) and orthotopic HCC implantation murine model (B) *P < 0.05, **P < 0.01, *** P< 0.001 versus the nontreated group.
Figure 8
Figure 8
The overall regulatory network involved in the inhibitory effect of ZJP on HCC.

Similar articles

See all similar articles

Cited by 1 article

References

    1. Baud V., Karin M. (2009). Is NF-kappaB a good target for cancer therapy? Hopes and pitfalls. Nat. Rev. Drug Discov. 8 (1), 33–40. 10.1038/nrd2781 - DOI - PMC - PubMed
    1. Calvisi D. F., Ladu S., Gorden A., Farina M., Conner E. A., Lee J. S., et al. (2006). Ubiquitous activation of ras and jak/stat pathways in human HCC. Gastroenterology 130 (4), 1117–1128. 10.1053/j.gastro.2006.01.006 - DOI - PubMed
    1. Chen Y., Kern T. S., Kiser P. D., Palczewski K. (2016). Eyes on systems pharmacology. Pharmacol. Res. 114, 39–41. 10.1016/j.phrs.2016.09.026 - DOI - PMC - PubMed
    1. Chen Y., Wei J., Zhang Y., Sun W., Li Z., Wang Q., et al. (2018). Anti-endometriosis mechanism of jiawei foshou san based on network pharmacology. Front. Pharmacol. 9, 811. 10.3389/fphar.2018.00811 - DOI - PMC - PubMed
    1. Chou S. T., Hsiang C. Y., Lo H. Y., Huang H. F., Lai M. T., Hsieh C. L., et al. (2017). Exploration of anti-cancer effects and mechanisms of Zuo-Jin-Wan and its alkaloid components in vitro and in orthotopic HepG2 xenograft immunocompetent mice. BMC Complement Altern. Med. 17 (1), 121. 10.1186/s12906-017-1586-6 - DOI - PMC - PubMed
Feedback