Compound-composed Chinese medicine of Huachansu triggers apoptosis of gastric cancer cells through increase of reactive oxygen species levels and suppression of proteasome activities

• Published in: Phytomedicine (Stuttgart) 2024-01, Vol.123, p.155169-155169, Article 155169
• Main Authors: Deng, Yi-Qing, Gao, Min, Lu, Dong, Liu, Qiu-Ping, Zhang, Run-Jing, Ye, Ji, Zhao, Jing, Feng, Zhi-Hui, Li, Qi-Zhang, Zhang, Hong
• Format: Article
• Language: English
• Subjects: Amphibian Venoms; Apoptosis; Cell Line, Tumor; Compound-composed Chinese medicine; gastric cancer; Humans; Medicine, Chinese Traditional; methodology; Phosphatidylinositol 3-Kinases - metabolism; proteasome; Proteasome Endopeptidase Complex; Proteomics; Proto-Oncogene Proteins c-akt - metabolism; reactive oxygen species; Reactive Oxygen Species - metabolism; Stomach Neoplasms - drug therapy; Stomach Neoplasms - metabolism
• ISSN: 0944-7113; 1618-095X
• DOI: 10.1016/j.phymed.2023.155169

•A new strategy was proposed for research of the compound-composed Chinese medicine based on network pharmacology and serum chemistry.•Compound-composed Chinese medicine of HCS possesses better apoptosis induction effects in gastric cancer cells than HCS.•Compound-composed Chinese medicine of HCS triggers apoptosis of gastric cancer cells majorly through increasing ROS levels and suppressing proteasome activities. Huachansu (HCS), a known Chinese patent drug extracted from the Chinese toad skin, is frequently used for the treatment of various advanced cancers, especially gastric cancer, due to the good therapeutic effect. However, it is rather difficult to clarify the active substances and molecular mechanisms involved owing to the lack of appropriate research strategies. We recently proposed the concept and research ideas of compound-composed Chinese medicine formula. To discover compound-composed Chinese medicine from Huachansu and to explore its mechanism of action in inducing apoptosis of gastric cancer cells. Network pharmacology combined with serum pharmacochemistry was utilized to screen the predominant active constituents from HCS against gastric cancer. Then, the compound-composed Chinese medicine of HCS (CCMH) was prepared according to their relative contents in serum. The pharmacological effects and potential mechanisms for CCMH were investigated by assays for cell viability, cell cycle, apoptosis, mitochondrial membrane potential (MMP), proteomics, reactive oxygen species (ROS), N-Acetylcysteine (NAC) antagonism, proteasome activity, and western blot. : CCMH was comprised of arenobufagin (11.14%), bufalin (18.67%), bufotalin (7.33%), cinobufagin (16.67%), cinobufotalin (16.74%), gamabufotalin (8.45%), resibufogenin (12.03%), and telocinobufagin (8.97%). CCMH evidently induced proliferation inhibition, cell cycle arrest, apoptosis, and MMP collapse in gastric cancer cells, possessing the better activities than HCS. Proteomic analysis showed that CCMH influenced ROS pathway, ubiquitin proteasome system, and PI3K/Akt and MAPK signaling pathways. CCMH markedly enhanced intracellular ROS levels in gastric cancer cells, which was reversed by NAC. Accordingly, NAC antagonized the apoptosis-inducing effect of CCMH. Significantly decreased proteasome 20S activity by CCMH was observed in gastric cancer cells. CCMH also regulated the expression of key proteins in PI3K/Akt and MAPK signaling pathways. CCMH possesses more significant apoptotic induction eff