Research ArticleExperimental Studies
Open Access

Effect of Recombinant Methioninase Dose and Timing on the Selective and Precise Elimination of Cancer Cells from Co-Cultured Normal Cells and on Methionine-dependent Rescue of Cancer Cells

JINSOO KIM, QINGHONG HAN, SHUKUAN LI, BYUNG MO KANG, KOHEI MIZUTA, YOHEI ASANO, YUTA MIYASHI, MICHAEL BOUVET and ROBERT M. HOFFMAN
Anticancer Research January 2026, 46 (1) 143-151; DOI: https://doi.org/10.21873/anticanres.17930

Abstract

Background/Aim: Methionine addiction is a fundamental and general hallmark of cancer, known as the Hoffman effect. The aim of the present study was to target methionine addiction of cancer cells co-cultured with normal cells with various doses of recombinant methioninase (rMETase) to precisely eliminate cancer cells and to determine the timing of rescue of cancer cells by methionine supplementation.

Materials and Methods: HCT116 human colon cancer cells were co-cultured with Hs-27 diploid human normal fibroblasts in 12-well tissue-culture plates containing Dulbecco’s Modified Eagle’s Medium (DMEM) with 10% fetal bovine serum. rMETase was added to the co-cultures at 0.25, 0.5, or 1.0 U/ml. In attempts to rescue the cancer cells, normal DMEM which contains 0.2 mM of methionine, replaced DMEM containing rMETase on days −2, −4 or −6. Co-cultures were observed for cell viability and proliferation using phase-contrast microscopy.

Results: The efficacy of rMETase to eliminate cancer cells co-cultured with normal cells was dose-dependent. rMETase at 0.25 U/ml allowed large numbers of cancer cells to remain in the co-cultures by day 12. rMETase at 0.5 U/ml and 1.0 U/ml largely eliminated the cancer cells from the co-cultures by day 12. rMETase at 1.0 U/ml was slightly toxic to the normal cells. However, at each dose of rMETase, even at 1.0 U/ml, DMEM could rescue the cancer cells even when added on day 6.

Conclusion: The ability of rMETase to eliminate cancer cells co-cultured with normal cells is dose-dependent. Even at high effective doses of rMETase, the cancer cells could be rescued by methionine supplementation up to at least day-6, indicating the need for the continuous presence of rMETase to selectively inhibit the cancer cells.

Keywords:

Introduction

Methionine addiction, defined as the dependence of cancer cells on exogenous methionine despite their ability to synthesize normal or greater amounts of methionine endogenously, is a fundamental and general metabolic hallmark of cancer termed the Hoffman effect (1-5). Comparison of methionine- and glucose-based positron emission tomography (PET) imaging has demonstrated that the Hoffman effect of methionine addiction is stronger than the Warburg effect of glucose addiction, respectively (6). Therefore, methionine addiction is a promising target for cancer therapy.

Methionine addiction has been therapeutically targeted because cancers show specific vulnerability to methionine restriction (7-11). An effective approach to methionine restriction of cancer cells involves the use of recombinant methioninase (rMETase), an enzyme which degrades methionine (5).

An internally-controlled system to compare cancer-selective efficacy of a potential therapy is the co-culture of cancer and normal cells which we devised 40 years ago to show that methionine-restricted culture medium, plus chemotherapy, could selectively eliminate cancer cells from co-cultures with normal cells (12). Recently we showed that rMETase could selectively eliminate cancer cells from co-cultures with normal cells (13).

In the present study, we show the ability of rMETase to eliminate cancer cells co-cultured with normal cells is dose-dependent. We also show that even at high doses of rMETase, the cancer cells can be rescued by methionine-containing medium, therefore indicating the necessity of continuous maintenance of rMETase to selectively inhibit cancer-cell growth.

Materials and Methods

Cell culture. HCT116 human colon-cancer cells and Hs-27 human fibroblasts were obtained from the American Type Culture Collection (Manassas, VA, USA). The cells were cultured in Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 with GlutaMAX™ supplement (DMEM/F-12), containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37°C in a humidified incubator containing 5% CO2.

Recombinant methioninase production. rMETase was produced by AntiCancer Inc. (San Diego, CA, USA) with fermentation of recombinant Escherichia coli transformed with the methioninase gene from Pseudomonas putida. The purification process involved heat treatment at 60°C, polyethylene glycol precipitation, and diethylaminoethyl (DEAE) Sepharose fast-flow ion-exchange column chromatography, as previously described (14).

Co-culture of HCT116 colon-cancer cells with Hs-27 normal fibroblasts and treatment with rMETase and rescue with methionine. HCT116 colon-cancer cells and Hs-27 normal fibroblasts were seeded together in 12-well plates at 5×104 cells each (total 1×105 cells/well). Twenty-four hours after seeding, co-cultures were treated with varying concentrations of rMETase at 0.25, 0.5, or 1.0 U/ml. In attempts to rescue cancer cells, normal methionine-containing DMEM/F-12 replaced the rMETase containing DMEM/F-12 on days 2, 4, and 6. Cell growth and morphology were monitored with phase-contrast microscopy (IX71, Olympus Corporation, Tokyo, Japan) on days 0, 4, 8, and 12 after treatment. Representative microscopic fields were photographed. Cell viability was semi-quantitatively assessed from phase-contrast images. Experiments were performed in triplicate.

Results

Selective elimination of HCT116 colon-cancer cells in co-culture with Hs-27 normal fibroblasts depends on the rMETase concentration. Continuous rMETase treatment selectively inhibited cancer-cell viability in a dose- and time-dependent manner (Figures 1-3). At 0.25 U/ml rMETase, selective inhibition of cancer cells was less pronounced, with scattered proliferating cancer cells remaining on day 12. At 0.5 U/ml rMETase, cancer cells were progressively eliminated from the co-cultures leaving viable normal cells. At 1.0 U/ml rMETase, nearly complete loss of cancer cells was observed, whereas normal cells were continuously present by day 12.

Figure 1.
Figure 1.

Phase-contrast images of HCT116 human colon-cancer cells co-cultured with Hs27 normal human fibroblasts treated with rMETase (0.25 U/ml) for 0, 4, 8, and 12 days without methionine rescue or with methionine rescue with DMEM/F12 on day 2, day 4, and day 6. Images were captured using an Olympus IX71 inverted microscope at 100× total magnification. Representative HCT116 colon-cancer cells are indicated with white arrows and representative Hs-27 normal fibroblasts are indicated with black arrows. rMETase: Recombinant methioninase. Please see Materials and Methods for details.

Figure 2.
Figure 2.

Phase-contrast images of HCT116 human colon-cancer cells co-cultured with Hs27 normal human fibroblasts treated with rMETase (0.5 U/ml) for 0, 4, 8, and 12 days without methionine rescue or with methionine rescue with DMEM/F12 on day 2, day 4, and day 6. Images were captured using an Olympus IX71 inverted microscope at 100× total magnification. Representative HCT116 colon-cancer cells are indicated with white arrows and representative Hs-27 normal fibroblasts are indicated with black arrows. rMETase: Recombinant methioninase. Please see Materials and Methods for details.

Figure 3.
Figure 3.

Phase-contrast images of HCT116 human colon-cancer cells co-cultured with Hs27 normal human fibroblasts treated with rMETase (1.0 U/ml) for 0, 4, 8, and 12 days without methionine rescue or with methionine rescue with DMEM/F12 on day 2, day 4, and day 6. Images were captured using an Olympus IX71 inverted microscope at 100× total magnification. Representative HCT116 colon-cancer cells are indicated with white arrows and representative Hs-27 normal fibroblasts are indicated with black arrows. rMETase: Recombinant methioninase. Please see Materials and Methods for details.

Rescue with methionine of rMETase-treated HCT116 colon-cancer cells in co-culture with Hs-27 normal fibroblasts depends on rMETase dose and methionine rescue schedule. Rescue with DMEM/F-12 on day 2 restored cancer-cell proliferation at all concentrations of rMETase, with cancer cells overtaking the co-cultures by day 12 (Figures 1-3). Rescue on day 4 resulted in nearly full recovery of cancer cells at all concentrations of rMETase, but their number was reduced compared to day-2 rescue (Figures 1-3). Rescue on day 6 failed to restore the full number of cancer cells by day 12 (Figures 1-3). However, significant numbers of HCT116 cancer cells recovered with DMEM/F12 rescue at day 6, even at 0.5 U/ml and 1.0 U/ml rMETase concentrations (Figures 2 and 3). At each dose of rMETase, even at 1.0 U/ml, DMEM/F12 could rescue some cancer cells when added on day 6 (Figures 1-3).

In the co-cultures treated with 0.5 U/ml and rescued on day 6 with methionine, the cancer cells still decreased by day 8 and normal cells were healthy and proliferating. In the co- cultures treated with 1.0 U/ml and rescued on day 6, the cancer cells decreased by day 8 but so did the normal cells (Figure 4). Therefore, 1.0 U/ml rMETase is slightly toxic to normal cells.

Figure 4.
Figure 4.

Phase-contrast images of HCT116 human colon-cancer cells co-cultured with Hs27 normal human fibroblasts treated with rMETase (0.25, 0.5, or 1.0 U/ml) and methionine rescue with DMEM/F12 on day 6. Images were obtained on day 4, day 6, and day 8 using an Olympus IX71 inverted microscope at 100× total magnification. Representative HCT116 colon cancer cells are indicated with white arrows and representative Hs-27 normal fibroblasts are indicated with black arrows. rMETase: Recombinant methioninase. Please see Materials and Methods for details.

Discussion

An effective anti-cancer agent is the one that selectively targets a cancer -specific vulnerability. Methionine addiction is a general cancer-specific vulnerability. Cancer-specific vulnerabilities are proven in co-cultures of normal and cancer cells whereby a potential agent selectively eliminates the cancer cells, allowing the normal cells to live and proliferate.

We showed 40 years ago that methionine restriction, combined with chemotherapy could selectively eliminate cancer cells co-cultured with normal cells, which remained healthy and proliferating (12). Very recently we showed rMETase alone could selectively eliminate cancer cells co-cultured with normal cells (13).

The present study showed that the ability of rMETase to eliminate cancer cells co-cultured with normal cells is dose-dependent. The present study also showed the necessity of continuous maintenance of rMETase, even at a high dose of rMETase, to prevent methionine rescue of cancer cells.

Oral-rMETase (o-rMETase) is being used as a dietary supplement in patients with metastatic cancer (15-24). The present results emphasize the need for precise doses of rMETase and continuous use.

rMETase is effective because it targets the fundamental hallmark of cancer, methionine addiction (1-11, 25-45).

The Hoffman effect of methionine addiction is stronger than the Warburg effect of glucose addiction of cancer as shown by comparison of methionine-based and glucose-based PET imaging of cancer patients, respectively (6).

Acknowledgements

This article is dedicated to the memory of A.R. Moossa, MD; Sun Lee, MD; Professor Philip Miles; Richard W. Erbe, MD; Professor Milton Plesur; Professor Gordon H. Sato; Professor Li Jiaxi; Masaki Kitajima, MD; Shigeo Yagi, Ph.D.; Jack Geller, MD; Joseph R Bertino, MD; J.A.R. Mead, Ph.D.; Eugene P. Frenkel, MD; Professor I. J. Fidler; John Mendelsohn, MD; Professor Lev Bergelson; Professor Sheldon Penman; Professor John R. Raper; Professor J.D. Watson and Joseph Leighton, MD. The Robert M. Hoffman Foundation for Cancer Research provided funds for the present study.

Footnotes

  • Authors’ Contributions

    JK and RMH designed the study. QH and SL produced rMETase. JK conducted all experiments and wrote the article. RMH revised the article. BMK, KM, YA, YM and MB critically read the manuscript.

  • Conflicts of Interest

    All Authors have no conflicts of interest or financial ties to disclose related to this study.

  • Artificial Intelligence (AI) Disclosure

    No artificial intelligence (AI) tools, including large language models or machine learning software, were used in the preparation, analysis, or presentation of this manuscript.

  • Received October 3, 2025.
  • Revision received October 24, 2025.
  • Accepted October 27, 2025.

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

References

  1. ↵
    1. Hoffman RM,
    2. Erbe RW
    : High in vivo rates of methionine biosynthesis in transformed human and malignant rat cells auxotrophic for methionine. Proc Natl Acad Sci U S A 73(5): 1523-1527, 1976. DOI: 10.1073/pnas.73.5.1523
    OpenUrlAbstract/FREE Full Text
    1. Coalson DW,
    2. Mecham JO,
    3. Stern PH,
    4. Hoffman RM
    : Reduced availability of endogenously synthesized methionine for S-adenosylmethionine formation in methionine-dependent cancer cells. Proc Natl Acad Sci USA 79(14): 4248-4251, 1982. DOI: 10.1073/pnas.79.14.4248
    OpenUrlAbstract/FREE Full Text
    1. Stern PH,
    2. Mecham JO,
    3. Wallace CD,
    4. Hoffman RM
    : Reduced free-methionine in methionine-dependent SV40-transformed human fibroblasts synthesizing apparently normal amounts of methionine. J Cell Physiol 117(1): 9-14, 1983. DOI: 10.1002/jcp.1041170103
    OpenUrlCrossRefPubMed
    1. Judde JG,
    2. Ellis M,
    3. Frost P
    : Biochemical analysis of the role of transmethylation in the methionine dependence of tumor cells. Cancer Res 49(17): 4859-4865, 1989.
    OpenUrlAbstract/FREE Full Text
  5. ↵
    1. Abo Qoura L,
    2. Balakin KV,
    3. Hoffman RM,
    4. Pokrovsky VS
    : The potential of methioninase for cancer treatment. Biochim Biophys Acta Rev Cancer 1879(4): 189122, 2024. DOI: 10.1016/j.bbcan.2024.189122
    OpenUrlCrossRef
  6. ↵
    1. Sato M,
    2. Sato T,
    3. Hozumi C,
    4. Han Q,
    5. Mizuta K,
    6. Morinaga S,
    7. Kang BM,
    8. Kobayashi N,
    9. Ichikawa Y,
    10. Nakajima A,
    11. Hoffman RM
    : [11C] Methionine PET vs. [18F]Fluorodeoxyglucose PET whole-body imaging to determine the extent of methionine-addiction compared to glucose-addiction of primary and metastatic cancer of the trunk in patients. Anticancer Res 44(9): 3891-3898, 2024. DOI: 10.21873/anticanres.17216
    OpenUrlAbstract/FREE Full Text
  7. ↵
    1. Tisdale MJ
    : Effect of methionine deprivation on methylation and synthesis of macromolecules. Br J Cancer 42(1): 121-128, 1980. DOI: 10.1038/bjc.1980.210
    OpenUrlCrossRefPubMed
    1. Stern PH,
    2. Hoffman RM
    : Elevated overall rates of transmethylation in cell lines from diverse human tumors. In Vitro 20(8): 663-670, 1984. DOI: 10.1007/BF02619617
    OpenUrlCrossRefPubMed
    1. Wang Z,
    2. Yip LY,
    3. Lee JHJ,
    4. Wu Z,
    5. Chew HY,
    6. Chong PKW,
    7. Teo CC,
    8. Ang HY,
    9. Peh KLE,
    10. Yuan J,
    11. Ma S,
    12. Choo LSK,
    13. Basri N,
    14. Jiang X,
    15. Yu Q,
    16. Hillmer AM,
    17. Lim WT,
    18. Lim TKH,
    19. Takano A,
    20. Tan EH,
    21. Tan DSW,
    22. Ho YS,
    23. Lim B,
    24. Tam WL
    : Methionine is a metabolic dependency of tumor-initiating cells. Nat Med 25(5): 825-837, 2019. DOI: 10.1038/s41591-019-0423-5
    OpenUrlCrossRefPubMed
    1. Yamamoto J,
    2. Han Q,
    3. Inubushi S,
    4. Sugisawa N,
    5. Hamada K,
    6. Nishino H,
    7. Miyake K,
    8. Kumamoto T,
    9. Matsuyama R,
    10. Bouvet M,
    11. Endo I,
    12. Hoffman RM
    : Histone methylation status of H3K4me3 and H3K9me3 under methionine restriction is unstable in methionine-addicted cancer cells, but stable in normal cells. Biochem Biophys Res Commun 533(4): 1034-1038, 2020. DOI: 10.1016/j.bbrc.2020.09.108
    OpenUrlCrossRefPubMed
  11. ↵
    1. Raboni S,
    2. Montalbano S,
    3. Stransky S,
    4. Garcia BA,
    5. Buschini A,
    6. Bettati S,
    7. Sidoli S,
    8. Mozzarelli A
    : A key silencing histone mark on chromatin is lost when colorectal adenocarcinoma cells are depleted of methionine by methionine γ-Lyase. Front Mol Biosci 8: 735303, 2021. DOI: 10.3389/fmolb.2021.735303
    OpenUrlCrossRefPubMed
  12. ↵
    1. Stern PH,
    2. Hoffman RM
    : Enhanced in vitro selective toxicity of chemotherapeutic agents for human cancer cells based on a metabolic defect. J Natl Cancer Inst 76(4): 629-639, 1986. DOI: 10.1093/jnci/76.4.629
    OpenUrlCrossRefPubMed
  13. ↵
    1. Kang BM,
    2. Han Q,
    3. Li S,
    4. Kim JS,
    5. Mizuta K,
    6. Asano Y,
    7. Miyashi Y,
    8. Bouvet M,
    9. Hoffman RM
    : Recombinant methioninase selectively eliminates cancer cells co-cultured with normal fibroblasts indicating the high-precision efficacy of targeting methionine addiction of cancer. Anticancer Res 45(10): 4193-4200, 2025. DOI: 10.21873/anticanres.17771
    OpenUrlAbstract/FREE Full Text
  14. ↵
    1. Tan Y,
    2. Xu M,
    3. Tan X,
    4. Tan X,
    5. Wang X,
    6. Saikawa Y,
    7. Nagahama T,
    8. Sun X,
    9. Lenz M,
    10. Hoffman RM
    : Overexpression and large-scale production of recombinantl-methionine-α-deamino-γ-mercaptomethane-lyase for novel anticancer therapy. Protein Expr Purif 9(2): 233-245, 1997. DOI: 10.1006/prep.1996.0700
    OpenUrlCrossRefPubMed
  15. ↵
    1. Han Q,
    2. Hoffman RM
    : Lowering and stabilizing PSA levels in advanced-prostate cancer patients with oral methioninase. Anticancer Res 41(4): 1921-1926, 2021. DOI: 10.21873/anticanres.14958
    OpenUrlAbstract/FREE Full Text
    1. Han Q,
    2. Hoffman RM
    : Chronic treatment of an advanced prostate-cancer patient with oral methioninase resulted in long-term stabilization of rapidly rising PSA levels. In Vivo 35(4): 2171-2176, 2021. DOI: 10.21873/invivo.12488
    OpenUrlAbstract/FREE Full Text
    1. Han Q,
    2. Tan Y,
    3. Hoffman RM
    : Oral dosing of recombinant methioninase is associated with a 70% drop in PSA in a patient with bone-metastatic prostate cancer and 50% reduction in circulating methionine in a high-stage ovarian cancer patient. Anticancer Res 40(5): 2813-2819, 2020. DOI: 10.21873/anticanres.14254
    OpenUrlAbstract/FREE Full Text
    1. Kubota Y,
    2. Han Q,
    3. Hamada K,
    4. Aoki Y,
    5. Masaki N,
    6. Obara K,
    7. Tsunoda T,
    8. Hoffman RM
    : Long-term stable disease in a rectal-cancer patient treated by methionine restriction with oral recombinant methioninase and a low-methionine diet. Anticancer Res 42(8): 3857-3861, 2022. DOI: 10.21873/anticanres.15877
    OpenUrlAbstract/FREE Full Text
    1. Kubota Y,
    2. Han Q,
    3. Masaki N,
    4. Hozumi C,
    5. Hamada K,
    6. Aoki Y,
    7. Obara K,
    8. Tsunoda T,
    9. Hoffman RM
    : Elimination of axillary-lymph-node metastases in a patient with invasive lobular breast cancer treated by first-line neo-adjuvant chemotherapy combined with methionine restriction. Anticancer Res 42(12): 5819-5823, 2022. DOI: 10.21873/anticanres.16089
    OpenUrlAbstract/FREE Full Text
    1. Kubota Y,
    2. Han Q,
    3. Morinaga S,
    4. Tsunoda T,
    5. Hoffman RM
    : Rapid reduction of CEA and stable metastasis in an NRAS-mutant rectal-cancer patient treated with FOLFIRI and bevacizumab combined with oral recombinant methioninase and a low-methionine diet upon metastatic recurrence after FOLFIRI and bevacizumab treatment alone. In Vivo 37(5): 2134-2138, 2023. DOI: 10.21873/invivo.13310
    OpenUrlAbstract/FREE Full Text
    1. Sato M,
    2. Han Q,
    3. Hozumi C,
    4. Kujiraoka H,
    5. Mizuta K,
    6. Morinaga S,
    7. Kang BM,
    8. Kobayashi N,
    9. Ichikawa Y,
    10. Nakajima A,
    11. Hoffman RM
    : First-line chemotherapy in combination with oral recombinant methioninase and a low-methionine diet for a stage IV inoperable pancreatic-cancer patient resulted in 40% tumor reduction and an 86% CA19-9 biomarker decrease. Anticancer Res 44(9): 3885-3889, 2024. DOI: 10.21873/anticanres.17215
    OpenUrlAbstract/FREE Full Text
    1. Sato M,
    2. Han Q,
    3. Mori R,
    4. Mizuta K,
    5. Kang BM,
    6. Morinaga S,
    7. Kobayashi N,
    8. Ichikawa Y,
    9. Nakajima A,
    10. Hoffman RM
    : Reduction of tumor biomarkers from very high to normal and extensive metastatic lesions to undetectability in a patient with stage IV HER2-positive breast cancer treated with low-dose trastuzumab deruxtecan in combination with oral recombinant methioninase and a low-methionine diet. Anticancer Res 44(4): 1499-1504, 2024. DOI: 10.21873/anticanres.16946
    OpenUrlAbstract/FREE Full Text
    1. Morinaga S,
    2. Han Q,
    3. Mizuta K,
    4. Kang BM,
    5. Yamamoto N,
    6. Hayashi K,
    7. Kimura H,
    8. Miwa S,
    9. Igarashi K,
    10. Higuchi T,
    11. Tsuchiya H,
    12. Demura S,
    13. Hoffman RM
    : Prostate cancer patient with lymph-node metastasis treated only with methionine restriction has stable disease for two years demonstrated with PET/CT and PSMA-PET scanning and PSA testing. Cancer Diagn Progn 5(1): 27-31, 2025. DOI: 10.21873/cdp.10408
    OpenUrlCrossRefPubMed
  24. ↵
    1. Morinaga S,
    2. Han Q,
    3. Mizuta K,
    4. Kang BM,
    5. Yamamoto N,
    6. Hayashi K,
    7. Kimura H,
    8. Miwa S,
    9. Igarashi K,
    10. Higuchi T,
    11. Tsuchiya H,
    12. Demura S,
    13. Hoffman RM
    : Complete response (CR) in a previously-progressing chronic lymphocytic leukemia (CLL) patient treated with methionine restriction in combination with first-line chemotherapy. Cancer Diagn Progn 5(1): 21-26, 2025. DOI: 10.21873/cdp.10407
    OpenUrlCrossRefPubMed
  25. ↵
    1. Sugimura T,
    2. Birnbaum SM,
    3. Winitz M,
    4. Greenstein JP
    : Quantitative nutritional studies with water-soluble, chemically defined diets. VIII. The forced feeding of diets each lacking in one essential amino acid. Arch Biochem Biophys 81(2): 448-455, 1959. DOI: 10.1016/0003-9861(59)90225-5
    OpenUrlCrossRefPubMed
    1. Halpern BC,
    2. Clark BR,
    3. Hardy DN,
    4. Halpern RM,
    5. 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 USA 71(4): 1133-1136, 1974. DOI: 10.1073/pnas.71.4.1133
    OpenUrlAbstract/FREE Full Text
    1. Hoffman RM,
    2. Jacobsen SJ,
    3. Erbe RW
    : Reversion to methionine independence in simian virus 40-transformed human and malignant rat fibroblasts is associated with altered ploidy and altered properties of transformation. Proc Natl Acad Sci USA 76(3): 1313-1317, 1979. DOI: 10.1073/pnas.76.3.1313
    OpenUrlAbstract/FREE Full Text
    1. Yamamoto J,
    2. Inubushi S,
    3. Han Q,
    4. Tashiro Y,
    5. Sugisawa N,
    6. Hamada K,
    7. Aoki Y,
    8. Miyake K,
    9. Matsuyama R,
    10. Bouvet M,
    11. Clarke SG,
    12. Endo I,
    13. Hoffman RM
    : Linkage of methionine addiction, histone lysine hypermethylation, and malignancy. iScience 25(4): 104162, 2022. DOI: 10.1016/j.isci.2022.104162
    OpenUrlCrossRefPubMed
    1. Chello PL,
    2. Bertino JR
    : Dependence of 5-methyltetrahydrofolate utilization by L5178Y murine leukemia cells in vitro on the presence of hydroxycobalamin and transcobalamin II. Cancer Res 33(8): 1898-1904, 1973.
    OpenUrlAbstract/FREE Full Text
    1. Aoki Y,
    2. Han Q,
    3. Tome Y,
    4. Yamamoto J,
    5. Kubota Y,
    6. Masaki N,
    7. Obara K,
    8. Hamada K,
    9. Wang JD,
    10. Inubushi S,
    11. Bouvet M,
    12. Clarke SG,
    13. Nishida K,
    14. Hoffman RM
    : Reversion of methionine addiction of osteosarcoma cells to methionine independence results in loss of malignancy, modulation of the epithelial-mesenchymal phenotype and alteration of histone-H3 lysine-methylation. Front Oncol 12: 1009548, 2022. DOI: 10.3389/fonc.2022.1009548
    OpenUrlCrossRefPubMed
    1. Yamamoto J,
    2. Aoki Y,
    3. Inubushi S,
    4. Han Q,
    5. Hamada K,
    6. Tashiro Y,
    7. Miyake K,
    8. Matsuyama R,
    9. Bouvet M,
    10. Clarke SG,
    11. Endo I,
    12. Hoffman RM
    : Extent and instability of trimethylation of histone H3 lysine increases with degree of malignancy and methionine addiction. Cancer Genomics Proteomics 19(1): 12-18, 2022. DOI: 10.21873/cgp.20299
    OpenUrlAbstract/FREE Full Text
    1. Sullivan MR,
    2. Darnell AM,
    3. Reilly MF,
    4. Kunchok T,
    5. Joesch-Cohen L,
    6. Rosenberg D,
    7. Ali A,
    8. Rees MG,
    9. Roth JA,
    10. Lewis CA,
    11. Vander Heiden MG
    : Methionine synthase is essential for cancer cell proliferation in physiological folate environments. Nat Metab 3(11): 1500-1511, 2021. DOI: 10.1038/s42255-021-00486-5
    OpenUrlCrossRef
    1. Ghergurovich JM,
    2. Xu X,
    3. Wang JZ,
    4. Yang L,
    5. Ryseck RP,
    6. Wang L,
    7. Rabinowitz JD
    : Methionine synthase supports tumour tetrahydrofolate pools. Nat Metab 3(11): 1512-1520, 2021. DOI: 10.1038/s42255-021-00465-w
    OpenUrlCrossRefPubMed
    1. Lin DW,
    2. Carranza FG,
    3. Borrego S,
    4. Lauinger L,
    5. Dantas de Paula L,
    6. Pulipelli HR,
    7. Andronicos A,
    8. Hertel KJ,
    9. Kaiser P
    : Nutrient control of splice site selection contributes to methionine addiction of cancer. Mol Metab 93: 102103, 2025. DOI: 10.1016/j.molmet.2025.102103
    OpenUrlCrossRefPubMed
    1. Andronicos A,
    2. Yoneda KC,
    3. Lin D-W,
    4. Law FV,
    5. Bae H,
    6. Basirattalab A,
    7. Graham NA,
    8. Jang C,
    9. Kaiser P
    : Carboxy-methylation of the catalytic subunit of protein phosphatase 2A (PP2Ac) integrates methionine availability with methionine addicted cancer cell proliferation. Biomolecules 15(9): 1210, 2025. DOI: 10.3390/biom15091210
    OpenUrlCrossRefPubMed
    1. Kaiser P
    : Methionine dependence of cancer. Biomolecules 10(4): 568, 2020. DOI: 10.3390/biom10040568
    OpenUrlCrossRefPubMed
    1. Mecham JO,
    2. Rowitch D,
    3. Wallace CD,
    4. Stern PH,
    5. Hoffman RM
    : The metabolic defect of methionine dependence occurs frequently in human tumor cell lines. Biochem Biophys Res Comm 117(2): 429-434, 1983. DOI: 10.1016/0006-291X(83)91218-4
    OpenUrlCrossRefPubMed
    1. Tisdale MJ
    : Effect of methionine deprivation on methylation and synthesis of macromolecules. Br J Cancer 42(1): 121-128, 1980. DOI: 10.1038/bjc.1980.210
    OpenUrlCrossRefPubMed
    1. Breillout F,
    2. Antoine E,
    3. Poupon MF
    : Methionine dependency of malignant tumors: a possible approach for therapy. J Natl Cancer Inst 82(20): 1628-1632, 1990. DOI: 10.1093/jnci/82.20.1628
    OpenUrlCrossRefPubMed
    1. Tan Y,
    2. Xu M,
    3. Hoffman RM
    : Broad selective efficacy of recombinant methioninase and polyethylene glycol-modified recombinant methioninase on cancer cells in vitro. Anticancer Res 30(4): 1041-1046, 2010.
    OpenUrlAbstract/FREE Full Text
    1. Gao X,
    2. Sanderson SM,
    3. Dai Z,
    4. Reid MA,
    5. Cooper DE,
    6. Lu M,
    7. Richie JP Jr.,
    8. Ciccarella A,
    9. Calcagnotto A,
    10. Mikhael PG,
    11. Mentch SJ,
    12. Liu J,
    13. Ables G,
    14. Kirsch DG,
    15. Hsu DS,
    16. Nichenametla SN and
    17. Locasale JW
    : Dietary methionine influences therapy in mouse cancer models and alters human metabolism. Nature 572(7769): 397-401, 2019. PMID: 31367041. DOI: 10.1038/s41586-019-1437-3
    OpenUrlCrossRefPubMed
    1. Stern PH,
    2. Wallace CD,
    3. Hoffman RM
    : Altered methionine metabolism occurs in all members of a set of diverse human tumor cell lines. J Cell Physiol 119(1): 29-34, 1984. DOI: 10.1002/jcp.1041190106
    OpenUrlCrossRefPubMed
    1. Hoffman RM
    : Altered methionine metabolism, DNA methylation and oncogene expression in carcinogenesis. A review and synthesis. Biochim Biophys Acta 738(1-2): 49-87, 1984. DOI: 10.1016/0304-419x(84)90019-2
    OpenUrlCrossRefPubMed
    1. Kawaguchi K,
    2. Miyake K,
    3. Han Q,
    4. Li S,
    5. Tan Y,
    6. Igarashi K,
    7. Kiyuna T,
    8. Miyake M,
    9. Higuchi T,
    10. Oshiro H,
    11. Zhang Z,
    12. Razmjooei S,
    13. Wangsiricharoen S,
    14. Bouvet M,
    15. Singh SR,
    16. Unno M,
    17. Hoffman RM
    : Oral recombinant methioninase (o-rMETase) is superior to injectable rMETase and overcomes acquired gemcitabine resistance in pancreatic cancer. Cancer Lett 432: 251-259, 2018. DOI: 10.1016/j.canlet.2018.06.016
    OpenUrlCrossRefPubMed
  45. ↵
    1. Yamamoto J,
    2. Aoki Y,
    3. Han Q,
    4. Sugisawa N,
    5. Sun Y,
    6. Hamada K,
    7. Nishino H,
    8. Inubushi S,
    9. Miyake K,
    10. Matsuyama R,
    11. Bouvet M,
    12. Endo I,
    13. Hoffman RM
    : Reversion from methionine addiction to methionine independence results in loss of tumorigenic potential of highly-malignant lung-cancer cells. Anticancer Res 41(2): 641-643, 2021. DOI: 10.21873/anticanres.14815
    OpenUrlAbstract/FREE Full Text
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Effect of Recombinant Methioninase Dose and Timing on the Selective and Precise Elimination of Cancer Cells from Co-Cultured Normal Cells and on Methionine-dependent Rescue of Cancer Cells
JINSOO KIM, QINGHONG HAN, SHUKUAN LI, BYUNG MO KANG, KOHEI MIZUTA, YOHEI ASANO, YUTA MIYASHI, MICHAEL BOUVET, ROBERT M. HOFFMAN
Anticancer Research Jan 2026, 46 (1) 143-151; DOI: 10.21873/anticanres.17930
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