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Thymol Enhances 5-Fluorouracil Cytotoxicity by Reducing Migration and Increasing Apoptosis and Cell Cycle Arrest in Esophageal Cancer Cells: An In-vitro Study

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Abstract

Esophageal cancer (EC) is a common cancer globally and has a low survival rate because it is often diagnosed at a late-stage, and is frequently chemotherapy resistant. Thymol is found in the herb thyme and has been reported to have potential anticancer properties. We aimed to explore the effects of thymol on the efficacy of 5-Fluorouracil (5-FU) treatment in an in vitro model of EC. We evaluated the synergistic effects of thymol with 5-FU using cell viability, apoptosis, cell cycle analysis, scratch wound healing assay, gelatin zymography, as well as reactive oxygen species (ROS) generation aproaches. The results indicated that thymol significantly decreased the viability of KYSE-30 cells in a time and dose-dependently. However, thymol had a minimal cytotoxic effect against normal fibroblasts. In addition, thymol significantly increased the chemosensitivity of EC KYSE-30 cells to 5-FU by promoting early and late apoptosis, inducing Sub-G1 and G2/M cell cycle arrest, and increasing ROS generation. Combination therapy of thymol with 5-FU also resulted in an increased expression of p53 and Bax, decreased expression of Bcl2, reduced activity of MMP-2, and reduced cell migration compared to single therapies. These findings suggest that thymol has the potential to promote the anticancer potency of 5-FU chemotherapy and inhibit metastasis. However, further studies using animal models are necessary to validate these results.

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Acknowledgements

We appreciate the assistance of the Clinical Research Development Unit of Akbar Hospital, Mashhad in conducting this research.We also would like to thank the Clinical Research Development Unit, Imam Reza Hospital, Mashhad University of Medical Sciences, for their assistance in this manuscript.

Funding

This study is supported by Mashhad University of Medical Sciences, Mashhad, Iran (grant code: 4021071).

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Author notes

  1. Ghazaleh Pouyamanesh and Nima Ameli are equal contributors as the first authors.

Authors and Affiliations

  1. Department of Medical Laboratory Sciences, Mashhad Branch, Islamic Azad University, Mashhad, Iran

    Ghazaleh Pouyamanesh

  2. Sinus and Surgical Endoscopic Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Nima Ameli

  3. School of Health and Social Care, Swansea University, Wales, UK

    Nima Ameli

  4. Faculty of Medicine, Zahedan University of Medical Sciences, Zahedan, Sistan and Baluchestan Province, Iran

    Yekta Metanat

  5. Medical School, Mashhad University of Medical Sciences, Mashhad, Iran

    Azar Khorrami

  6. Department of Medical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

    Farzaneh Abbasinezhad-Moud

  7. Endovascular Section, Neurosurgical Department, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran

    Farid Qoorchi Moheb Seraj

  8. Department of Medical Education, Brighton & Sussex Medical School, Falmer, Brighton, Sussex, BN1 9PH, UK

    Gordon A. Ferns

  9. Clinical Research Development Unit, Imam Reza Hospital, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

    Afsane Bahrami

  10. Clinical Research Development Unit of Akbar Hospital, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

    Afsane Bahrami

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  1. Ghazaleh Pouyamanesh
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Contributions

A.B, G.P, N.A, Y.M, A.Kh, F.A-M, and F.Q.M.S: Methodology, Investigation, Formal analysis, Conceptualization, Writing—Original Draft. G.A.F and A.B: Supervision, Funding acquisition, Validation, Resources, Project administration, Writing—Review & Editing.

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Correspondence to Afsane Bahrami.

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Pouyamanesh, G., Ameli, N., Metanat, Y. et al. Thymol Enhances 5-Fluorouracil Cytotoxicity by Reducing Migration and Increasing Apoptosis and Cell Cycle Arrest in Esophageal Cancer Cells: An In-vitro Study. Ind J Clin Biochem (2024). https://doi.org/10.1007/s12291-024-01219-7

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