Original Articles

L-Methioninase as a Selective Anticancer Agent: Dose-Dependent Cytotoxicity and Metastasis Suppression in Methionine-Dependent Tumor Cells

Abstract

L-methioninase (L-Met), a methionine-degrading enzyme, has shown potential for anticancer therapy. Many tumor tissues have a limited ability to produce methionine and depend on external sources; hence, these tumors can be targeted by methionine-based treatments. The present study was conducted to investigate the effects of L-Met on cancer cells, particularly hepatocellular carcinoma (HepG2) and pancreatic carcinoma (PANC-1), and to evaluate its viability as a therapeutic agent. Various techniques, including ammonium sulfate precipitation, dialysis, ion-exchange chromatography, and gel filtration chromatography, were employed to purify the enzyme L-Met. A cytotoxicity test was conducted against HepG2 and PANC-1 cells (at 25-200 µg/mL concentrations), using the MTT to evaluate cell viability, total nuclear intensity (TNI), and cell membrane permeability (CMP). Statistical analysis was done using one-way ANOVA and Dunnett's multiple comparisons test to compare study groups. L-Met displayed dose-dependent growth inhibition of the specified cell lines. The PANC-1 cell line exhibited an IC50 of 64.68 µg/mL, indicating a higher sensitivity to L-Met compared to WRL 68 normal cells, which had an IC50 of 214.0 µg/mL. Regarding HepG2, an even lower IC₅₀ of 66.44 µg/mL was observed, further confirming the selective targeting of cancer cells by L-Met. Treatment with L-Met at a 200 µg/mL concentration significantly decreased TNI and CMP levels in both the PANC-1 and HepG2 cell lines, indicating increased cytotoxicity and compromised membrane integrity. Additionally, L-Met reduced matrix metalloproteinase activities in both cancer cell lines, a critical factor in metastasis. Our study demonstrates the dose-dependent cytotoxic effects of L-Met on methionine-dependent tumor cells, specifically HepG2 and PANC-1. These findings highlight the need for optimized L-Met dosing strategies in cancer treatment, particularly for methionine-dependent malignancies, paving the way for its potential use in targeted cancer therapy.

1. Javia BM, Gadhvi MS, Vyas SJ, Ghelani A, Wirajana N, Dudhagara DR. A review on L-methioninase in cancer therapy: Precision targeting, advancements and diverse applications for a promising future. Int J Biol Macromol 2024:130997.
2. Bu T, Lan J, Jo I, Zhang J, Bai X, He S, et al. Structural Basis of the Inhibition of L-Methionine γ-Lyase from Fusobacterium nucleatum. Int J Mol Sci 2023;24:1651.
3. Wise DR, Thompson CB. Glutamine addiction: a new therapeutic target in cancer. Trends Biochem Sci 2010;35:427-33.
4. Halpern BC, Clark BR, Hardy DN, Halpern RM, Smith RA. The effect of replacement of methionine by homocystine on survival of malignant and normal adult mammalian cells in culture. Proc Natl Acad Sci 1974;71:1133-6.
5. Cellarier E, Durando X, Vasson M, Farges M, Demiden A, Maurizis J, et al. Methionine dependency and cancer treatment. Cancer Treat Rev 2003;29:489-99.
6. Kubota Y, Han Q, Aoki Y, Masaki N, Obara K, Hamada K, et al. Synergy of combining methionine restriction and chemotherapy: the disruptive next generation of cancer treatment. Cancer Diagn Progn 2023;3:272.
7. Tan Y, Xu M, Hoffman RM. Broad selective efficacy of recombinant methioninase and polyethylene glycol-modified recombinant methioninase on cancer cells in vitro. Anticancer Res 2010;30:1041-6.
8. Kawaguchi K, Igarashi K, Li S, Han Q, Tan Y, Miyake K, et al. Recombinant methioninase (rMETase) is an effective therapeutic for BRAF-V600E-negative as well as-positive melanoma in patient-derived orthotopic xenograft (PDOX) mouse models. Oncotarget 2018;9:915.
9. Sundar WA, Nellaiah H. Production of methioninase from Serratia marcescens isolated from soil and its anti-cancer activity against Dalton’s Lymphoma Ascitic (DLA) and Ehrlich Ascitic Carcinoma (EAC) in swiss albino mice. Trop J Pharm Res 2013;12:699-704.
10. Sorin M, Watkins D, Gilfix BM, Rosenblatt DS. Methionine dependence in tumor cells: The potential role of cobalamin and MMACHC. Mol Genet Metab 2021;132:155-61.
11. Pokrovsky VS, Abo Qoura L, Demidova EA, Han Q, Hoffman RM. Targeting methionine addiction of cancer cells with methioninase. Biochemistry 2023;88:944-52.
12. Dayanand K, Nadumane VK. Effect of physicochemical parameters on the L-methioninase activity of Methylobacterium sp. and its in vitro anticancer activity in combination with tamoxifen citrate. Futur J Pharm Sci 2023;9:94.
13. Zou Y, Yuan Y, Zhou Q, Yue Z, Liu J, Fan L, et al. The Role of Methionine Restriction in Gastric Cancer: A Summary of Mechanisms and a Discussion on Tumor Heterogeneity. Biomolecules 2024;14:161.
14. Zhao T, Lum JJ. Methionine cycle-dependent regulation of T cells in cancer immunity. Front Oncol 2022;12:969563.
15. Sedillo JC, Cryns VL. Targeting the methionine addiction of cancer. Am J Cancer Res 2022;12:2249.
16. Aldawood AS, Al-Ezzy RM. Cytotoxicity of L-Methioninase Purified from Clinical Isolates of Pseudomonas Species in Cancer Cell Lines. Al-Rafidain J Med Sci 2024;6:46-9.
17. Qoura LA, Balakin KV, Hoffman RM, Pokrovsky VS. The potential of methioninase for cancer treatment. BBA Reviews on Cancer 2024:189122.
18. Muharram MM. Recombinant Engineering of L-Methioninase Using Two Different Promoter and Expression Systems and in vitro Analysis of Its Anticancer Efficacy on Different Human Cancer Cell Lines. Pak J Biol Sci 2016;19:106-14.
19. Hoffman R. Altered methionine metabolism and transmethylation in cancer. Anticancer Res 1985;5:1-30.
20. El-Sayed AS. Microbial L-methioninase: production, molecular characterization, and therapeutic applications. Appl Microbiol Biotechnol 2010;86:445-67.
21. Davis CD, Uthus EO. DNA methylation, cancer susceptibility, and nutrient interactions. Exp Biol Med 2004;229:988-95.
22. Eymard N, Bessonov N, Volpert V, Kurbatova P, Gueyffier F, Nony P. Pharmacokinetic/pharmacodynamic model of a methionine starvation based anti-cancer drug. Med Biol Eng Comput 2023;61:1697-722.
23. Guo H-Y, Herrera H, Groce A, Hoffman RM. Expression of the biochemical defect of methionine dependence in fresh patient tumors in primary histoculture. Cancer Res 1993;53:2479-83.
24. Kokkinakis DM, Schold Jr SC, Hori H, Nobori T. Effect of long‐term depletion of plasma methionine on the growth and survival of human brain tumor xenografts in athymic mice. Nutr Cancer 1997;29:195-204.
25. Zhao Y, Butler EB, Tan M. Targeting cellular metabolism to improve cancer therapeutics. Cell Death Dis 2013;4:e532-e.
26. Calvo MB, Figueroa A, Pulido EG, Campelo RG, Aparicio LA. Potential role of sugar transporters in cancer and their relationship with anticancer therapy. Int J Endocrinol 2010;2010:205357.
27. Ishikawa N, Oguri T, Isobe T, Fujitaka K, Kohno N. SGLT gene expression in primary lung cancers and their metastatic lesions. Jpn J Cancer Res 2001;92:874-9.
28. Younes M, Lechago LV, Somoano JR, Mosharaf M, Lechago J. Wide expression of the human erythrocyte glucose transporter Glut1 in human cancers. Cancer Res 1996;56:1164-7.
29. McCracken AN, Edinger AL. Nutrient transporters: the Achilles’ heel of anabolism. Trends in Endocrinology & Metabolism 2013;24:200-8.
30. Ito S, Fukusato T, Nemoto T, Sekihara H, Seyama Y, Kubota S. Coexpression of glucose transporter 1 and matrix metalloproteinase-2 in human cancers. J Natl Cancer Inst 2002;94:1080-91.
31. Hassanein M, Hoeksema MD, Shiota M, Qian J, Harris BK, Chen H, et al. SLC1A5 mediates glutamine transport required for lung cancer cell growth and survival. Clin Cancer Res 2013;19:560-70.
32. Macheda ML, Rogers S, Best JD. Molecular and cellular regulation of glucose transporter (GLUT) proteins in cancer. J Cell Physiol 2005;202:654-62.
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IssueVol 63 No 2 (2025) QRcode
SectionOriginal Articles
Keywords
L-methioninase Pseudomonas aeruginosa Hepatocellular carcinoma Pancreatic carcinoma

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How to Cite
1.
Noor Al-Owaidi AA, Abdullah Jebor M. L-Methioninase as a Selective Anticancer Agent: Dose-Dependent Cytotoxicity and Metastasis Suppression in Methionine-Dependent Tumor Cells. Acta Med Iran. 2025;63(2):83-97.