Identification of potential anti-tumor targets and mechanisms of HuaChanSu injection using network pharmacology and cytological experiments in Breast cancer

PLoS One. 2024 May 16;19(5):e0303650. doi: 10.1371/journal.pone.0303650. eCollection 2024.

Abstract

HuaChanSu (HCS) or Cinobufacini injection is an aqueous extract of the dried skin of Bufo bufo gargarigans, and has anti-tumor effects. The aim of this study was to evaluate the possible therapeutic effect of HCS against breast cancer (BRCA) using cytology, network pharmacology, and molecular biology approaches. The half-inhibitory concentration (IC50) of HCS in the BRCA cells was determined by cytotoxicity assay, and were accordingly treated with high and low doses HCS in the TUNEL and scratch assays. The potential targets of HCS in the BRCA cells were identified through functional enrichment analysis and protein-protein interaction (PPI) networks, and verified by molecular docking. The expression levels of key signaling pathways-related proteins in HCS-treated BRCA cells by western blotting. HCS inhibited the proliferation and migration of MCF-7 and MDA-MB-231 cells, and induced apoptosis in a dose-dependent manner. Furthermore, we screened 289 core HCS targets against BRCA, which were primarily enriched in the PI3K-AKT, MAPK chemokines, and other. signaling pathways. In addition, PIK3CA, PIK3CD, and MTOR were confirmed as HCS targets by molecular docking. Consistent with this, we observed a reduction in the expression levels of phosphorylated PI3K, AKT, and MTOR in the HCS-treated BRCA cells. Taken together, our findings suggest that HCS inhibits the growth of BRCA cells by targeting the PI3K-AKT pathway, and warrants further investigation as a therapeutic agent for treating patients with BRCA.

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • Apoptosis* / drug effects
  • Breast Neoplasms* / drug therapy
  • Breast Neoplasms* / metabolism
  • Breast Neoplasms* / pathology
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cell Proliferation* / drug effects
  • Female
  • Humans
  • MCF-7 Cells
  • Molecular Docking Simulation
  • Network Pharmacology*
  • Protein Interaction Maps / drug effects
  • Proto-Oncogene Proteins c-akt / metabolism
  • Signal Transduction* / drug effects

Grants and funding

This work is supported by the Youth Program Research Project of the Affiliated Traditional Chinese Medicine Hospital of Guangzhou Medical University (No. 20210131), Guangzhou Chinese Medicine and Integrative Medicine Science and Technology Project (No. 20232A011005), and Scientific Research Project of Guangdong Provincial Administration of Traditional Chinese Medicine (No. 20221324 and No. 20210078). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. There was no additional external funding received for this study.