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Article

Two faces of Amitriptyline in an in vitro study on C6 glioma cells: The effects of Amitriptyline and its combination with Temozolomide and radiation

  • Authors:
    • Anna M. Bielecka-Wajdman
    • Miłosz Gołyszny
    • Wojciech Majewski
    • Tomasz Cichoń
    • Krzysztof Szczepanik
    • Ewa Obuchowicz
  • View Affiliations / Copyright

    Affiliations: Department of Pharmacology, Medical University of Silesia, 40‑752 Katowice, Poland, Department of Radiotherapy, Gliwice branch, Maria Sklodowska‑Curie National Research Institute of Oncology, 44‑101 Gliwice, Poland, Center of Translational Research and Molecular Biology of Cancer, Gliwice branch, Maria Sklodowska‑Curie National Research Institute of Oncology, 44‑101 Gliwice, Poland
  • Article Number: 38
    |
    Published online on: March 30, 2026
       https://doi.org/10.3892/mco.2026.2947
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Abstract

Failures in treating glioma have made it necessary to search for new therapies, especially those that have immunological properties. Amitriptyline (AMI) is not only used in the treatment of major depression but is also effective in the therapy for neuropathic and chronic cancer pain. Previous data have indicated anticancer and anti‑inflammatory effects of AMI. However, at present, its effect on glioma cells in combination with the standard treatment, namely temozolomide (TEMO, the first‑line cytostatic) and radiotherapy, has not been investigated. It would also be interesting to determine whether the mechanism of the immunomodulatory action of AMI is associated with its effect on the programmed death ligand‑1 (PD‑L1) expression, a key target for anticancer therapies. In the present study, the effect of AMI or its concomitant use with AMI and TEMO and/or radiation (10 Gy) on the viability, mortality (MTT, Trypan blue), proliferation (BrdU), colony forming (microscopic analysis) of C6 glioma cells and PD‑L1 expression (enzyme immunoassay) was investigated. Although AMI induced the anticancer effects, it attenuated the effects of radiation. In radiated cell cultures, the combination of AMI and TEMO provoked the formation of larger glioma cell colonies and reversed the cellular effects of radiation. Moreover, AMI suppressed the expression of PD‑L1 in cells that had been exposed to or had not been exposed to radiation, whereas radiation enhanced its expression. Because AMI exhibited promising anticancer properties including an interesting, previously unknown immunomodulatory effect, it appears that its potential therapeutic should be verified in an in vivo study.
View Figures

Figure 1

Effects of AMI (10 µM) and /or TEMO
(1 mM) (or radiation) administration into culture medium on (A)
viability of cells - MTT assay (%); (B) mortality of the cells
analyzed in EVE cell counter (%); (C) proliferation of the cells
(BrdU positive) (%), (D) PD-L1 expression (ng/ml). Experiments were
conducted on C6 rat glioma cells. Data are presented as mean ± SEM
from three independent experiments (n=9 per group). The results
were analyzed using two-way ANOVA followed by post hoc Bonferroni's
test. ***P<0.001 vs. Glioma (non-rad);
****P<0.0001 vs. Glioma (non-rad);
&&&&P<0.0001 vs. Glioma (rad);
##P<0.01 and ####P<0.0001 vs. Glioma
(non-rad) + AMI; ^^^^P<0.0001 vs. Glioma (non-rad) +
AMI, $$$$P<0.0001 vs. Glioma (non-rad) + AMI + TEMO.
AMI, amitriptyline; TEMO, temozolomide.

Figure 2

Representative pictures C6 glioma
cells: (A) untreated; (B) exposed to 72 h of amitriptyline (10 µM);
(C) TEMO; (D) AMI + TEMO combination; (E-H) or radiation.
Observations were performed using JuliStage (NanoEntek) in a bright
field (9 views for each group; magnification, x40).
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Spandidos Publications style
Bielecka-Wajdman AM, Gołyszny M, Majewski W, Cichoń T, Szczepanik K and Obuchowicz E: Two faces of Amitriptyline in an <em>in vitro</em> study on C6 glioma cells: The effects of Amitriptyline and its combination with Temozolomide and radiation. Mol Clin Oncol 24: 38, 2026.
APA
Bielecka-Wajdman, A.M., Gołyszny, M., Majewski, W., Cichoń, T., Szczepanik, K., & Obuchowicz, E. (2026). Two faces of Amitriptyline in an <em>in vitro</em> study on C6 glioma cells: The effects of Amitriptyline and its combination with Temozolomide and radiation. Molecular and Clinical Oncology, 24, 38. https://doi.org/10.3892/mco.2026.2947
MLA
Bielecka-Wajdman, A. M., Gołyszny, M., Majewski, W., Cichoń, T., Szczepanik, K., Obuchowicz, E."Two faces of Amitriptyline in an <em>in vitro</em> study on C6 glioma cells: The effects of Amitriptyline and its combination with Temozolomide and radiation". Molecular and Clinical Oncology 24.5 (2026): 38.
Chicago
Bielecka-Wajdman, A. M., Gołyszny, M., Majewski, W., Cichoń, T., Szczepanik, K., Obuchowicz, E."Two faces of Amitriptyline in an <em>in vitro</em> study on C6 glioma cells: The effects of Amitriptyline and its combination with Temozolomide and radiation". Molecular and Clinical Oncology 24, no. 5 (2026): 38. https://doi.org/10.3892/mco.2026.2947
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Spandidos Publications style
Bielecka-Wajdman AM, Gołyszny M, Majewski W, Cichoń T, Szczepanik K and Obuchowicz E: Two faces of Amitriptyline in an <em>in vitro</em> study on C6 glioma cells: The effects of Amitriptyline and its combination with Temozolomide and radiation. Mol Clin Oncol 24: 38, 2026.
APA
Bielecka-Wajdman, A.M., Gołyszny, M., Majewski, W., Cichoń, T., Szczepanik, K., & Obuchowicz, E. (2026). Two faces of Amitriptyline in an <em>in vitro</em> study on C6 glioma cells: The effects of Amitriptyline and its combination with Temozolomide and radiation. Molecular and Clinical Oncology, 24, 38. https://doi.org/10.3892/mco.2026.2947
MLA
Bielecka-Wajdman, A. M., Gołyszny, M., Majewski, W., Cichoń, T., Szczepanik, K., Obuchowicz, E."Two faces of Amitriptyline in an <em>in vitro</em> study on C6 glioma cells: The effects of Amitriptyline and its combination with Temozolomide and radiation". Molecular and Clinical Oncology 24.5 (2026): 38.
Chicago
Bielecka-Wajdman, A. M., Gołyszny, M., Majewski, W., Cichoń, T., Szczepanik, K., Obuchowicz, E."Two faces of Amitriptyline in an <em>in vitro</em> study on C6 glioma cells: The effects of Amitriptyline and its combination with Temozolomide and radiation". Molecular and Clinical Oncology 24, no. 5 (2026): 38. https://doi.org/10.3892/mco.2026.2947
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