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Piperlongumine and bortezomib synergically inhibit cholangiocarcinoma via ER stress–induced cell death

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Abstract

Cholangiocarcinoma (CCA) is a lethal malignancy of the cholangiocytes lining the biliary tree. Only 25% of affected patients are eligible for resection due to late-stage diagnosis. Systemic chemotherapy is recommended for those inoperable patients; however, an inadequate response to such treatment remains a significant obstacle. Piperlongumine (PL) is a biologically active alkaloid that selectively kills various cancer cells through the induction of reactive oxygen species (ROS). The role of PL has been shown through its inhibiting the ubiquitin–proteasome system. The mechanism of PL-induced CCA cell death was investigated by inhibiting the UPS and testing the therapeutic potential of combining PL and the proteasome inhibitor bortezomib. A single treatment with PL or BTZ suppressed CCA cell growth. Combined treatment with PL with BTZ produced a synergistic interaction, evidenced by (1) a combination index of < 1 and (2) induction of cell cycle arrest and down-regulation of cell cycle markers. PL induced the accumulation of poly-ubiquitinated proteins in CCA cells but did not affect proteasome activity. PL, in combination with BTZ, amplified the accumulation of poly-ubiquitinated proteins in CCA cells, leading to an endoplasmic reticulum (ER) stress response through the induction of X-box binding protein mRNA splicing. Moreover, PL-combined BTZ promoted the activation of a proapoptotic unfolded protein response via the ATF4-CHOP axis. PL induced CCA cell death via increased accumulation of the poly-ubiquitinated proteins. PL also enhanced the anti-cancer activity of BTZ via ER stress–induced CCA cell death. Thus, the combination of PL and BTZ has potential as an alternative therapeutic option for CCA.

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All data generated or analyzed during the present study are included in this published article.

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Acknowledgements

We thank Mr. Bryan Roderick Hamman for assistance with the English-language presentation of the manuscript.

Funding

This work was supported by the Young Research Grants, Thailand Research Fund (TRG5680041) for C. Talabnin and the Development and Promotion of Science and Technology Talents Project (DPST) for funding and scholarship to Narathip Naradun.

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Authors and Affiliations

  1. School of Chemistry, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand

    Narathip Naradun & Chutima Talabnin

  2. School of Pathology, Institute of Medicine, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand

    Krajang Talabnin

  3. Biochemistry Laboratory Unit, The Center for Scientific and Technology Equipment, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand

    Kanyavee Issarangkul na Ayuttha

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  1. Narathip Naradun
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  2. Krajang Talabnin
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  4. Chutima Talabnin
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Contributions

CT was the grant holder. CT conceived and designed the present study. NN and CT performed the experiments. CT, NN, and KT analyzed the data. KT and KIA provided guardian advice and resources. NN drafted the manuscript. CT revised the manuscript. All authors read and approved the published version of the manuscript.

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Correspondence to Chutima Talabnin.

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Naradun, N., Talabnin, K., Ayuttha, K.I.n. et al. Piperlongumine and bortezomib synergically inhibit cholangiocarcinoma via ER stress–induced cell death. Naunyn-Schmiedeberg's Arch Pharmacol 396, 109–120 (2023). https://doi.org/10.1007/s00210-022-02305-4

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