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Phenformin inhibits proliferation, invasion, and angiogenesis of cholangiocarcinoma cells via AMPK-mTOR and HIF-1A pathways

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Abstract

Phenformin (Phen), a potent activator of AMPK, is effective against some resistant cancers. This study evaluated the inhibition of proliferation, migration, invasion, and angiogenesis by Phen in aggressive cancer cells and investigated the underlying mechanism of the inhibition. Cholangiocarcinoma (CCA) KKU-156 and KKU-452 cells were used in this study. The results showed that Phen suppressed cell proliferation and induced apoptosis in both cells. Phen suppressed migration and invasion of cancer cells in wound healing and transwell chamber assays, respectively. The effects were associated with depletions of glutathione (GSH) and decreased glutathione redox ratio which represents cellular redox state. The redox stress was linked with the loss of mitochondrial transmembrane potential, as evaluated by JC-1 assay. The effect of Phen on angiogenesis was performed using HUVEC cultured cells. Phen alone did not affect tube formation of HUVEC cells. However, conditioned media from CCA cell cultures treated with Phen suppressed the tube-like structure formation. The antitumor effect of Phen was associated with AMPK activation and suppression of mTOR phosphorylation, HIF-1A, and VEGF protein expression. In conclusion, Phen inhibits cell proliferation, migration, invasion, and angiogenesis probably through AMPK-mTOR and HIF-1A-VEGF pathways. Phen may be repurposed as chemoprevention of cancer.

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Funding

This work was supported by research grant from National Research Council of Thailand through Khon Kaen University (No. 61003302) and grant-in-aid from Faculty of Medicine (IN62302), Khon Kaen University, Thailand. RJ was supported by Studentship from Cholangiocarcinoma Research Institute, Khon Kaen University. We acknowledge Dr. Justin Thomas Reese for editing the manuscript via Publication Clinic KKU.

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Authors and Affiliations

  1. Department of Pharmacology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    Rattanaporn Jaidee, Sarinya Kongpetch, Laddawan Senggunprai, Auemduan Prawan & Veerapol Kukongviriyapan

  2. Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, 40002, Thailand

    Sarinya Kongpetch, Laddawan Senggunprai, Auemduan Prawan & Veerapol Kukongviriyapan

  3. Department of Physiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    Upa Kukongviriyapan

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  1. Rattanaporn Jaidee
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  2. Sarinya Kongpetch
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  3. Laddawan Senggunprai
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  4. Auemduan Prawan
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  5. Upa Kukongviriyapan
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  6. Veerapol Kukongviriyapan
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Contributions

Conceived and designed the study: VK. Performed the experiments: RJ, LS, SA, AP, and VK. Provided study materials: SA. LS, and AP. Analyzed the data: RJ, SA, LS, AP, and UK. Wrote the manuscript: VK and RJ.

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Correspondence to Veerapol Kukongviriyapan.

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Jaidee, R., Kongpetch, S., Senggunprai, L. et al. Phenformin inhibits proliferation, invasion, and angiogenesis of cholangiocarcinoma cells via AMPK-mTOR and HIF-1A pathways. Naunyn-Schmiedeberg's Arch Pharmacol 393, 1681–1690 (2020). https://doi.org/10.1007/s00210-020-01885-3

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  • DOI: https://doi.org/10.1007/s00210-020-01885-3

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