In vitro synergistic cytotoxic activity of oral hypoglycemic agent (Metformin) with Methotrexate against cancerous cells

Authors

  • Fatima Rizvi Dow International Medical College, Karachi, Pakistan

DOI:

https://doi.org/10.35845/abms.2025.1.397

Keywords:

Metformin, MTT assay, cytotoxic, synergistic, CDI index

Abstract

OBJECTIVE: Evaluation of synergistic cytotoxic effects of add-on therapy of Metformin with Methotrexate in different cancerous cells.

METHODOLOGY: This interventional study was undertaken at BMSI, JPMC, in conjunction with PCMD. The trial lasted three months. To appraise the Cytotoxic activity of metformin only and in combination with methotrexate, we employed cells from breast cancer (MCF-7 and MDA-MB-231), vaginal cancer (Hela), and colorectal adenocarcinoma (HT-29 cell line). We employed the most appropriate MTT tests to assess cytotoxic effects.

RESULTS:   When the percentage viabilities of the examined cell lines were compared, it was discovered that combination therapies of Metformin and Methotrexate dramatically lowered the percentage viabilities and had synergistic cytotoxic effects. There was a significant difference in the percent viability of cells representing breast cancer (MCF-7, MDA-MB-231), colorectal carcinoma (HT-29), and vaginal carcinoma representative cells (Hela cell lines) as assessed by MTT assay between Metformin alone and Metformin with Methotrexate. CDI values of each cell line for combination therapy were 0.702 ± 0.034, 0.67 ± 0.019, 0.69 ± 0.019, and 0.73 ± 0.040 for MDA-231, Hela, MCF-7, and HT-29 cells, respectively.

CONCLUSION: This research showed that Metformin also has synergistic effects with Methotrexate, in addition to having antiproliferative effects on the studied cancer cell lines.

References

Brown JS, Amend SR, Austin RH, Gatenby RA, Hammarlund EU, Pienta KJ. Updating the Definition of Cancer. Mol Cancer Res. 2023; 21(11):1142-1147.

Anand P, Kunnumakkara AB, Sundaram C, Harikumar KB, Tharakan ST, Lai OS, Sung B, Aggarwal BB. Cancer is a preventable disease that requires major lifestyle changes. Pharm Res. 2008; 25(9):2097-116.

Amin ARMR, Karpowicz PA, Carey TE, Arbiser J, Nahta R, Chen ZG, Dong JT, Kucuk O, Khan GN, Huang GS et al. Evasion of anti-growth signaling: A key step in tumorigenesis and potential target for treatment and prophylaxis by natural compounds. Semin Cancer Biol. 2015; 35 Suppl: S55-S77.

Shlyakhtina Y, Moran KL, Portal MM. Genetic and Non-Genetic Mechanisms Underlying Cancer Evolution. Cancers. 2021; 13(6):1380.

Nagai H, Kim YH. Cancer prevention from the perspective of global cancer burden patterns. J Thorac Dis. 2017; 9(3):448-451.

Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021; 71(3):209-249.

Xu CX, Zhu HH, Zhu YM. Diabetes and cancer: Associations, mechanisms, and implications for medical practice. World J Diabetes. 2014; 5(3):372-80.

Nasri H, Rafieian-Kopaei M. Metformin: Current knowledge. J Res Med Sci. 2014; 19(7):658-64.

Kim HJ, Lee S, Chun KH, Jeon JY, Han SJ, Kim DJ, Kim YS, Woo JT, Nam MS, Baik SH, Ahn KJ, Lee KW. Metformin reduces the risk of cancer in patients with type 2 diabetes: An analysis based on the Korean National Diabetes Program Cohort. Medicine (Baltimore). 2018; 97(8):e0036.

Lange C, Machado Weber A, Schmidt R, Schroeder C, Strowitzki T, Germeyer A. Changes in protein expression due to metformin treatment and hyperinsulinemia in a human endometrial cancer cell line. PLoS One. 2021; 16(3):e0248103.

Papadakos SP, Argyrou A, Lekakis V, Arvanitakis K, Kalisperati P, Stergiou IE, Konstantinidis I, Schizas D, Koufakis T, Germanidis G, Theocharis S. Metformin in Esophageal Carcinoma: Exploring Molecular Mechanisms and Therapeutic Insights. Int J Mol Sci. 2024; 25(5):2978.

Saraei P, Asadi I, Kakar MA, Moradi-Kor N. The beneficial effects of metformin on cancer prevention and therapy: a comprehensive review of recent advances. Cancer Manag Res. 2019; 11:3295-3313.

Duarte D, Vale N. Evaluation of synergism in drug combinations and reference models for future orientations in oncology. Curr Res Pharmacol Drug Discov. 2022; 3:100-110.

Bijnsdorp IV, Giovannetti E, Peters GJ. Analysis of drug interactions. Methods Mol Biol. 2011; 731:421-434.

Ali A, Manzoor MF, Ahmad N, Aadil RM, Qin H, Siddique R, Riaz S, Ahmad A, Korma SA, Khalid W, Aizhong L. The Burden of Cancer, Government Strategic Policies, and Challenges in Pakistan: A Comprehensive Review. Front Nutr. 2022; 9:940514.

Tufail, M., Wu, C. Exploring the Burden of Cancer in Pakistan: An Analysis of 2019 Data. J Epidemiol Glob Health 2023; 13: 333–343.

Pliszka M, Szablewski L. Associations between Diabetes Mellitus and Selected Cancers. Int J Mol Sci. 2024; 25(13):7476.

Roshan MH, Shing YK, Pace NP. Metformin as an adjuvant in breast cancer treatment. SAGE Open Med. 2019; 7:2050312119865114.

Amengual-Cladera E, Morla-Barcelo PM, Morán-Costoya A, Sastre-Serra J, Pons DG, Valle A, Roca P, Nadal-Serrano M. Metformin: From Diabetes to Cancer-Unveiling Molecular Mechanisms and Therapeutic Strategies. Biology (Basel). 2024;13(5):302.

Yang C, Zhao N, Li D, Zou G, Chen Y. Metformin improves the sensitivity of ovarian cancer cells to chemotherapeutic agents. Oncol Lett. 2019; 18(3):2404-2411.

Iliopoulos D, Hirsch HA, Struhl K. Metformin decreases the dose of chemotherapy for prolonging tumor remission in mouse xenografts involving multiple cancer cell types. Cancer Res. 2011; 71(9):3196-201.

Bekezhankyzy Z, Nurzhan S, Berdigaliyev N, Sergazy S, Maulenkul T, Aljofan M. The antiproliferative potential and mechanism of action of metformin in MCF-7 cells. Future Sci OA. 2023; 9(5):FSO859.

Galal MA, Al-Rimawi M, Hajeer A, Dahman H, Alouch S, Aljada A. Metformin: A Dual-Role Player in Cancer Treatment and Prevention. Int J Mol Sci. 2024; 25(7):4083.

Cejuela M, Martin-Castillo B, Menendez JA, Pernas S. Metformin and Breast Cancer: Where Are We Now? Int J Mol Sci. 2022; 23(5):2705.

Talib WH, Awajan D, Hamed RA, Azzam AO, Mahmod AI, Al-Yasari IH. Combination Anticancer Therapies Using Selected Phytochemicals. Molecules. 2022; 27(17):5452.

Published

2025-05-27

How to Cite

1.
Rizvi F. In vitro synergistic cytotoxic activity of oral hypoglycemic agent (Metformin) with Methotrexate against cancerous cells. Adv Basic Med Sci [Internet]. 2025May27 [cited 2025May30];9(1). Available from: https://abms.kmu.edu.pk/index.php/abms/article/view/397