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  3. Unveiling the Anticancer Potential and Molecular Mechanisms of Fangchinoline Against Cholangiocarcinoma Using FTIR Microspectroscopy, In Vitro and In Silico Approaches

Unveiling the Anticancer Potential and Molecular Mechanisms of Fangchinoline Against Cholangiocarcinoma Using FTIR Microspectroscopy, In Vitro and In Silico Approaches

  • Integr Cancer Ther. 2025 Jan-Dec:24:15347354251396513. doi: 10.1177/15347354251396513.
Piman Pocasap 1 2,  Karnchanok Kaimuangpak 1 2,  Krittaya Phukmee 1,  Auemduan Prawan 1 2,  Sarinya Kongpetch 1 2,  Laddawan Senggunprai 1 2
Affiliations

Affiliations

  • 1 Department of Pharmacology, Faculty of Medicine, Khon Kaen University, Thailand.
  • 2 Cholangiocarcinoma Research Institute, Khon Kaen University, Thailand.
PMID: 41261767 DOI: 10.1177/15347354251396513
Abstract

Objectives: The limited effectiveness of traditional chemotherapeutic agents against cholangiocarcinoma (CCA) highlights the need for novel compounds. This study aimed to investigate the Anticancer potential of fangchinoline (FCL) and its underlying mechanisms of action in CCA cells.

Methods: Biomolecular changes associated with FCL-induced cell death were determined by Fourier transform infrared spectroscopy. The apoptosis-inducing effects of FCL were investigated using annexin V/7-amino-actinomycin staining, caspase-9 activity, JC-1 transmembrane potential, and Reactive Oxygen Species (ROS) assays. Western blotting of total cell lysates was used to assess protein expression, normalized to β-actin. The Proteome Profiler™ Human Phospho-Kinase Array Kit was used to assess changes in the phosphorylation patterns of cell survival-associated proteins. Network pharmacology analysis was conducted to predict FCL targets. The interaction between the compound and its potential targets were demonstrated by molecular docking and verified by western blot analysis.

Results: FCL induced biochemical alterations in CCA cells. A series of in vitro analyses demonstrated that FCL induced mitochondria-associated apoptotic cell death in KKU-100 cells, with ROS induction contributing to this effect. Furthermore, FCL altered the phosphorylation status of several survival-related kinases, including Yes, c-Jun, ERK1/2, HSP27, and STAT5. Network pharmacology analysis identified Src as a central hub protein with the highest degree of connectivity among the signaling proteins affected by FCL. Consistently, western blot analysis confirmed a marked reduction in Src phosphorylation following FCL treatment. Molecular docking further supported this finding, showing that FCL interacted with the catalytic residues of Src.

Conclusion: The Anticancer potential of FCL against CCA was demonstrated using several approaches. These findings indicated a potential therapeutic opportunity for FCL in CCA.

Keywords

FTIR; Src; apoptosis; cholangiocarcinoma; fangchinoline.

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