Document Type : Research Articles
Authors
1
Department of Pharmacology, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand.
2
Medical Degree Program, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand.
3
Department of Physiology, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand.
4
Department of Pharmacology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
5
Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Thailand.
6
Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, Thailand.
7
Center for Advanced Studies in Nanotechnology for Chemical, Food and Agricultural Industries, Kasetsart University Institute for Advanced Studies, Kasetsart University, Bangkok, Thailand.
8
Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Songkhla, Thailand.
9
Phytomedicine and Pharmaceutical Biotechnology Excellence Center, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Songkhla, Thailand.
Abstract
Objective: To evaluate the anticancer mechanisms of Chamuangone against cholangiocarcinoma (CCA) cells. Methods: Chamuangone was tested for cytotoxicity against KKU-100 and KKU-452 cells for 24 and 48 h. Apoptosis, cell proliferation, mitochondrial membrane potential, and intracellular reactive oxygen species (ROS) levels were assessed using Annexin V, Ki-67, JC-1 assays, and DCFH-DA fluorescence probe, respectively. Oncology proteins expression was measured. Results: Chamuangone inhibited CCA cell growth in a dose- and time- dependent manner, with IC50 values in KKU- 100 cells of 1.175 and 0.331 μg/mL at 24 and 48 hours, respectively; in KKU- 452 cells, the IC50 values were 1.208 and 0.428 μg/mL. Consequently, Chamuangone at 1.5 and 3.0 μg/mL effectively induced both early and late apoptosis in a statistically significant manner, which correlated with a marked reduction in cell proliferation, as evidenced by the decrease in Ki-67 positive populations to 49.04% and 17.02%, respectively. Chamuangone at concentrations of 0.75, 1.5, and 3.0 μg/mL significantly induced mitochondrial dysfunction by reducing the Red/Green fluorescence ratio across all time points (3–24 h), indicating a loss of mitochondrial membrane potential that triggers apoptosis. The induction of intracellular oxidative stress was indicated by a significant increase in the high-dose group of Chamuangone. Moreover, it also suppressed the expression of key ROS- and oxidative stress–associated oncogenic proteins, including Carbonic Anhydrase IX, Enolase2, CXCL8/IL-8, Galectin-3, EGFR/ErbB1, Progranulin, FGF basic, Dkk-1, p27/kip1, Mesothelin, Survivin, leading to redox imbalance and apoptosis in KKU-100 cells. Conclusion: Chamuangone inhibits CCA cell proliferation by inducing apoptosis through mechanisms involving the suppression of the Ki67, loss of mitochondrial membrane potential, intracellular ROS accumulation, and downregulation of oncogenic-related proteins involved in proliferation, survival, angiogenesis, and oxidative stress. Thus, Chamuangone has significant potential as a lead compound for the development of novel CCA therapeutics.
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