Evaluation of L-Methioninase as a Targeted Anticancer Agent in Colorectal Cancer and Renal Cell Carcinoma
,
Abstract
The employment of L-Methioninase (L-Met), an enzyme degrading L-methionine, as a possible anticancer therapy has been provided that it selectively destroys cancer cells, which require methionine to grow. Many tumor tissues have a limited capacity for methionine production and depend on exogenous supplies of this amino acid. Therefore, they could be selectively attacked by methioninase-based treatment. The present study was carried out to investigate the effect of L-Met on cancer cells, especially colorectal cancer (HCT-116) and renal cell carcinoma (A498), and its potential as a therapeutic agent. Various techniques, such as ammonium sulfate precipitation, dialysis, ion-exchange chromatography, and gel filtration chromatography, were employed in this study for enzyme purification of L-Met. Cytotoxicity testing was performed against HCT-116 and A498 cells in the range of 25-200 µg/mL concentration by the MTT assay for viable cell quantification, Total Nuclear Intensity (TNI), and Cell Membrane Permeability (CMP). The statistical analysis was done using one-way ANOVA and Dunnett's multiple comparisons test to compare multiple groups. The optimal temperature for enzyme activity was 37° C, with pH 7 offering the best enzyme stability. Further investigation into incubation time revealed that a 48-hour period maximized enzyme yield. The enzyme was purified using a combination of ammonium sulfate precipitation, ion-exchange chromatography, and gel filtration, achieving a 4.6-fold increase in purity. Characterization of the purified enzyme revealed a molecular weight of approximately 55 kDa, with optimal activity at pH 7 and 37°C. The enzyme demonstrated strong potential for therapeutic applications, showing dose-dependent cytotoxicity against colorectal cancer (HCT-116) and renal cancer (A498) cell lines, with significant inhibitory effects at concentrations as low as 25 µg/mL for colorectal cancer cells. The study highlights the potential of P. aeruginosa-derived L-Met showed strong activity in colorectal cancer, while activity in renal cell carcinoma was lower or inconclusive.
2. Sharma R, Kannourakis G, Prithviraj P, Ahmed N. Precision medicine: an optimal approach to patient care in renal cell carcinoma. Front Med 2022;9:766869.
3. Jin H, Wang L, Bernards R. Rational combinations of targeted cancer therapies: background, advances and challenges. Nat Rev Drug Discov 2023;22:213-34.
4. Qoura LA, Balakin KV, Hoffman RM, Pokrovsky VS. The potential of methioninase for cancer treatment. Biochim Biophys Acta Rev Cancer 2024:189122.
5. Bin P, Wang C, Zhang H, Yan Y, Ren W. Targeting methionine metabolism in cancer: opportunities and challenges. Trends Pharmacol Sci 2024;45:395-405.
6. Sanderson SM, Gao X, Dai Z, Locasale JW. Methionine metabolism in health and cancer: a nexus of diet and precision medicine. Nat Rev Cancer 2019;19:625-37.
7. Maggi M, Scotti C. Enzymes in metabolic anticancer therapy. Therapeutic Enzymes: Function and Clinical Implications 2019:173-99.
8. Javia BM, Gadhvi MS, Vyas SJ, Ghelani A, Wirajana N, Dudhagara DR. A review on L-Met in cancer therapy: Precision targeting, advancements and diverse applications for a promising future. Int J Biol Macromol 2024:130997.
9. Mármol I, Sánchez-de-Diego C, Pradilla Dieste A, Cerrada E, Rodriguez Yoldi MJ. Colorectal carcinoma: a general overview and future perspectives in colorectal cancer. Int J Mol Sci 2017;18:197.
10. Mohd AB, Ghannam RA, Mohd OB, Elayan R, Albakri K, Huneiti N, et al. Etiologies, gross appearance, histopathological patterns, prognosis, and best treatments for subtypes of renal carcinoma: an educational review. Cureus 2022;14.
11. Chen J, Cui L, Lu S, Xu S. Amino acid metabolism in tumor biology and therapy. Cell Death & Disease 2024;15:42.
12. Gutiérrez-Salmerón M, Lucena SR, Chocarro-Calvo A, García-Martínez JM, Orozco RMM, García-Jiménez C. Metabolic and hormonal remodeling of colorectal cancer cell signalling by diabetes. Endocr Relat Cancer 2021;28:191-206.
13. Mazumder S, Higgins PJ, Samarakoon R. Downstream targets of VHL/HIF-α signaling in renal clear cell carcinoma progression: mechanisms and therapeutic relevance. Cancers 2023;15:1316.
14. Dhankhar R, Kawatra A, Mohanty A, Gulati P. Microbial enzymes used in prodrug activation for cancer therapy: insights and future perspectives. Current Protein and Peptide Science 2021;22:514-25.
15. Diala ES, Cheah MS, Rowitch D, Hoffman RM. Extent of DNA methylation in human tumor cells. J Natl Cancer Inst 1983;71:755-64.
16. Hoffman RM. Altered methionine metabolism, DNA methylation and oncogene expression in carcinogenesis: a review and synthesis. BBA Reviews on Cancer 1984;738:49-87.
17. Michalak EM, Burr ML, Bannister AJ, Dawson MA. The roles of DNA, RNA and histone methylation in ageing and cancer. Nat. Rev Mol Cell Biol 2019;20:573-89.
18. Erichsen L, Thimm C, Santourlidis S. Methyl group metabolism in differentiation, aging, and cancer. Int J Mol Sci 2022;23:8378.
19. 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.
20. Vande Voorde J, Steven RT, Najumudeen AK, Ford CA, Dexter A, Gonzalez-Fernandez A, et al. Metabolic profiling stratifies colorectal cancer and reveals adenosylhomocysteinase as a therapeutic target. Nat Metab 2023;5:1303-18.
21. Liu S, Locasale JW. Delineating a role for methionine metabolism in colorectal cancer. Cancer Res 2023;83:3833-4.
22. Wang X, Guo X, Yu W, Li C, Gui Y, Cai Z. Expression of methionine adenosyltransferase 2A in renal cell carcinomas and potential mechanism for kidney carcinogenesis. BMC cancer. 2014;14:1-9.
23. Garg S, Miousse IR. Rescue of Methionine Dependence by Cobalamin in a Human Colorectal Cancer Cell Line.
| Files | ||
| Issue | Vol 63 No 2 (2025) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/acta.v63i2.18961 | |
| Keywords | ||
| L-met Pseudomonas aeruginosa Colorectal cancer Renal cell carcinoma | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
