Research ArticleExperimental Studies
Open Access

Ultra-low Concentrations of Cisplatinum Down to the IC10 in Combination With Recombinant Methioninase Are Synergistically Effective Against Lung Cancer Cells In Vitro and In Vivo

YOHEI ASANO, QINGHONG HAN, SHUKUAN LI, BYUNG MO KANG, JIN SOO KIM, YUTA MIYASHI, MICHAEL BOUVET, NORIO YAMAMOTO, KATSUHIRO HAYASHI, HIROAKI KIMURA, SHINJI MIWA, KENTARO IGARASHI, TAKASHI HIGUCHI, SEI MORINAGA, HIROYUKI TSUCHIYA, SATORU DEMURA and ROBERT M. HOFFMAN
In Vivo March 2026, 40 (2) 803-812; DOI: https://doi.org/10.21873/invivo.14238

Abstract

Background/Aim: To determine whether methionine restriction using recombinant methioninase (rMETase) enhances the efficacy of ultra-low-dose cisplatinum against lung cancer cells in vitro, and whether combining a methionine-restricted (MR) diet with low-dose cisplatinum can inhibit lung cancer growth in vivo with reduced toxicity.

Materials and Methods: Human A549 lung adenocarcinoma cells were treated with rMETase and cisplatinum in vitro. Cell viability was assessed after 72 hours using the WST-8 reagent. The IC50 value of rMETase was determined, and synergy was evaluated by combining rMETase at its IC50 with cisplatinum at its determined IC10-IC50. For in vivo analysis, A549 xenografts were established in nude mice and assigned to four groups: control: standard-dose cisplatinum [6 mg/kg, intraperitoneally (i.p.), weekly]; low-dose cisplatinum (3 mg/kg, i.p., weekly) + a methionine-restricted (MR) diet; or the MR diet alone. Treatments were administered for two weeks, with tumor size and body weight were monitored.

Results: For A549 lung-cancer cells the IC50 value of rMETase was 0.64 U/ml. Combination treatment with rMETase (IC50) and cisplatinum (IC10-IC50) synergistically reduced cell viability compared with either agent alone, even at the IC10 of cisplatinum. In vivo, A549 tumor eradication was observed only in the low-dose cisplatinum + MR diet group. Standard-dose cisplatinum alone and MR-alone showed delayed or limited efficacy. Body-weight loss was minimal in the low-dose cisplatinum + MR group compared with the standard-dose cisplatinum group, indicating reduced systemic toxicity.

Conclusion: Methionine restriction enhances the efficacy of ultra-low-dose cisplatinum on lung cancer cells in vitro. Low-dose cisplatinum in combination with an MR diet prevented lung-cancer growth in nude mice. The present approach of cancer therapy may help reduce platinum-related toxicity and improve treatment outcomes, suggesting further investigation for clinical translation.

Keywords:

Introduction

Lung cancer remains the leading cause of cancer-related mortality worldwide, accounting for nearly 1.8 million deaths annually (1). Although molecularly-targeted therapies and immune-checkpoint inhibitors have improved outcomes in selected patient populations, platinum-based chemotherapy–particularly cisplatinum–continues to serve as first-line treatment for both small-cell and non-small-cell lung cancers (2, 3). However, cisplatinum efficacy is limited by severe dose-dependent toxicities, including nephrotoxicity and myelosuppression (4-6). Therefore, potential strategies that enhance cisplatinum efficacy while minimizing its required dose are of considerable clinical importance.

Methionine addiction is a fundamental and general hallmark of cancer, termed the Hoffman effect (7-22). Methionine restriction inhibits tumor growth and enhances sensitivity to cisplatinum of multiple malignancies, including lung cancer (15-19). Mechanistically, methionine restriction perturbs the abnormal transmethylation metabolism of cancer cells and induces cancer-specific late-S/G2-phase cell-cycle arrest, thereby potentiating the effects of DNA-damaging agents such as cisplatinum (15-20).

Preclinical studies have demonstrated that methionine restriction, including enzymatic depletion using recombinant methioninase (rMETase), acts synergistically with cisplatinum in osteosarcoma, bladder-cancer, and colon-cancer mouse models, enabling effective dose reduction (15-19). Nevertheless, whether methionine restriction can similarly potentiate cisplatinum efficacy while allowing dose reduction in lung cancer remains unknown.

The present study aimed to investigate whether methionine restriction enhances the efficacy of ultra-low-dose cisplatinum against lung-cancer cells in vitro and in vivo.

Materials and Methods

Cell culture and reagents. The human lung-adenocarcinoma cell line A549 was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin at 37°C in an incubator with 5% CO2. Cisplatinum was obtained from WG Critical Care, LLC (Paramus, NJ, USA).

Production of recombinant methioninase (rMETase). rMETase was produced at AntiCancer Inc (San Diego, CA, USA) by fermentation of recombinant E. coli, containing the Pseudomonas putida methioninase gene. Purification used a 60°C heat-step, polyethylene-glucol precipitation and DEAE Sepharose ion-exchange chomatography, as previously described (23).

Determination of IC50 of rMETase and IC10-50 of cisplatinum on A549 cells. A549 cells were cultured in 96-well plates at 1.0×103 cells/well in 100 μl of DMEM for 24 h. Cells were treated with rMETase (0.0625-8 U/ml) or cisplatinum (0.5-64 μM) for 72 h. After cell culture, 10 μl of the cell-viability reagent WST-8 (Dojindo Laboratories, Kumamoto, Japan) was then added to each well and incubated for 1 h. Subsequently, absorbance at 450 nm was measured using a microplate reader (Sunrise, Tecan, Männedorf, Switzerland). Drug-sensitivity curves were generated using Microsoft Excel for Mac 2024 (ver. 16.89.1), GraphPad Prism 10.4.1 (GraphPad Software, Inc., San Diego, CA, USA), and ImageJ (ver. 1.54g). The half maximal inhibitory concentration (IC50) value of rMETase and values of cisplatinum (from IC10 to IC50) were calculated based on drug-sensitivity curves.

Determination of the synergistic efficacy of the combination of rMETase and decreasing cisplatinum on A549 lung cancer cells. After culturing cells for 24 h in a 96-well plate as described above, cells were divided into the following eight groups and treated for 72 h: 1) untreated control (DMEM); 2) rMETase (IC50); 3) cisplatinum (IC50); 4) rMETase + cisplatinum (each at IC50); 5) rMETase (IC50) + cisplatinum (IC40); 6) rMETase (IC50) + cisplatinum (IC30); 7) rMETase (IC50) + cisplatinum (IC20); and 8) rMETase (IC50) + cisplatinum (IC10). After treatment, cell viability was determined as described above.

Mouse husbandry. In the present study, 5- to 6-week-old athymic nu/nu nude mice, produced by AntiCancer Inc. (San Diego, CA, USA), were used. The mice were housed in a barrier facility on a high-efficiency particulate arrestance (HEPA)-filtered rack under standard conditions of 12-hour light/dark cycles. All experiments were conducted according to protocols specifically approved for this study by the AntiCancer Institutional Animal Care and Use Committee. The procedures adhered to the principles and guidelines for the care and use of animals outlined by the National Institutes of Health (NIH). Each experiment was performed in accordance with the Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines version 2.0. To minimize pain for the mice, all experiments were performed under anesthesia. The anesthetic solution contained 20 mg/kg of ketamine, 15.2 mg/kg of xylazine, and 0.48 mg/kg of acepromazine maleate and was administered at a volume of 0.02 ml.

Study design for the determination of the efficacy of the combination of low-dose cisplatinum and rMETase on A549 lung cancer growing subcutaneously in nude mice. Cultured A549 cells (1.0×106 cells) were suspended in 100 μl of phosphate-buffered saline (PBS) and injected subcutaneously into the right flank of nude mice. Two weeks after injection, 20 mice with A549 subcutaneous tumors grown to 50-100 mm3 were selected and randomly assigned to four treatment groups (five mice per group) for a two-week treatment using a methionine-restricted (MR) diet (TD.90262; Inotiv, Inc., West Lafayette, IN, USA) or a normal diet (Inotiv): 1) untreated group (normal diet); 2) standard-dose cisplatinum [6 mg/kg intraperitoneal (i.p.) injection weekly] and a normal diet; 3) MR diet + low-dose cisplatinum (3 mg/kg i.p. weekly); 4) MR diet (Figure 1). Measurements of the tumor’s long and short axes using calipers and body-weight measurements were made twice a week. Tumor volume was calculated from measured tumor size using the following formula: tumor volume (mm3)=long axis (mm)×short axis (mm)×short axis (mm)×1/2.

Figure 1.
Figure 1.

Drug sensitivity curves of (A) recombinant methioninase (rMETase) and (B) cisplatinum (CDDP) on A549 lung-cancer cells in vitro. Please see Materials and Methods for details.

Statistical analysis. All data are presented as the mean ± standard error of the mean (SEM). Differences between groups were analyzed using a one-way analysis of variance (ANOVA), followed by Tukey’s multiple comparison test. All statistical analyses were conducted using EZR (Jichi Medical University, Saitama Medical Center, Japan). A p-value ≤0.05 was considered statistically significant. All in vitro experiments were carried out in triplicate and repeated independently twice. In vivo experiments had n=5.

Results

Efficacy of the IC10-50 of cisplatinum combined with the IC50 of rMETase on A549 lung cancer cells in vitro. Drug sensitivity curves were generated for rMETase and cisplatinum in A549 cells, and the inhibitory concentrations were calculated. For A549 lung cancer cells, the IC50 value for rMETase was 0.64 U/ml. For cisplatinum the IC10 was 0.16 μM; IC20 was 0.38 μM; IC30 was 0.67 μM; IC40 was 1.06 μM; and IC50 was 1.61 μM (Figure 1). The combination of rMETase at its IC50 concentration with cisplatinum at concentrations between IC10 and IC50 produced a synergistic decrease in cell viability compared to treatment with rMETase alone or cisplatinum alone at their IC50 (Figure 2).

Figure 2.
Figure 2.

Treatment of A549 lung-cancer cells with recombinant methioninase (rMETase) and cisplatinum at differenct concentration. The combination of rMETase and ultra-low concentrations of cisplatinum was highly effective against A549 cells. (*p-value <0.05). CDDP: Cisplatinum. Please see Materials and Methods for details.

Efficacy of low-dose cisplatinum combined with rMETase on subcutaneous A549 tumors in nude mice. Subcutaneous A549 tumors in nude mice were eradicated by low-dose cisplatinum combined with an MR diet (Please see Figure 3 for schema and Figure 4 for results). The tumor volume in the high-dose-cisplatinum-alone-treated mice decreased more than tumors in mice on the MR diet alone, but the difference was not statistically significant (Figure 4). Neither treatment significantly inhibited tumor growth compared to the untreated normal-diet group (Figure 4). Body weight in the control and MR diet groups increased steadily during treatment, with a greater increase observed in the control group than in the MR diet group (Figure 5). Both the standard-dose cisplatinum and low-dose-cisplatinum + MR-diet groups had a decrease in body weight immediately after cisplatinum administration, followed by recovery. At the final measurement, body weight in the low-dose cisplatinum + MR diet group was nearly identical to that in the MR diet group, whereas the standard-dose cisplatinum group had the greatest body weight loss, although the difference was not statistically significant. Only the standard-dose-cisplatinum group showed a final body weight lower than the pre-treatment weight (Figure 6).

Figure 3.
Figure 3.

Treatment of A549 tumors in nude mice with rMETase and low- and high-cisplatinum doses. Group 1 (untreated control): vehicle [PBS, intraperitoneal (i.p.) injection, weekly] with a normal diet. Group 2 (CDDP): standard-dose cisplatinum (6 mg/kg, i.p., weekly) with a normal diet. Group 3 (Low-CDDP + MR): low-dose CDDP (3 mg/kg, i.p., weekly) combined with a methionine-restricted (MR) diet. Group 4 (MR): MR diet only. Tumor volume and body weight were measured twice per week. CDDP: Cisplatinum. MR: Methionine restricted. Please see Materials and Methods for details.

Figure 4.
Figure 4.

Efficacy of an MR diet alone; high-dose cisplatinum alone; or low-dose cisplatinum plus an MR diet on subcutaneous A549 tumors in nude mice. Control: vehicle phosphate-buffered saline (PBS), intraperitoneal (i.p.) injection weekly] with a normal diet; CDDP: standard-dose cisplatinum (6 mg/kg, i.p., weekly) with a normal diet; Low-CDDP + MR: low-dose cisplatinum (3 mg/kg, i.p., weekly) combined with an MR diet; MR: methionine-restricted diet; CDDP: cisplatinum. (*p-value <0.05). Please see Materials and Methods for details.

Figure 5.
Figure 5.

Body weight changes of nude mice in after treatment with high-dose cisplatinum; an MR diet; or low-dose cisplatinum plus an MR diet. Control: vehicle phosphate-buffered saline (PBS), intraperitoneal (i.p.) injection weekly] with a normal diet; CDDP: standard-dose cisplatinum (6 mg/kg, i.p., weekly) with a normal diet; Low-CDDP + MR: low-dose cisplatinum (3 mg/kg, i.p., weekly) combined with an MR diet; MR: methionine restricted diet; CDDP: cisplatinum. Please see Materials and Methods for details.

Figure 6.
Figure 6.

Comparison of body weight in nude mice before and after the treatment period with high-dose cisplatinum; an MR diet plus low-dose cisplatinum or an MR diet. Control: vehicle phosphate-buffered saline (PBS) intraperitoneal (i.p.) injection weekly] with a normal diet; CDDP: standard-dose cisplatinum (6 mg/kg, i.p., weekly) with a normal diet; Low-CDDP + MR: low-dose cisplatinum (3 mg/kg, i.p., weekly) combined with an MR diet. MR: Methionine restricted diet; CDDP: cisplatinum. Please see Materials and Methods for details.

Discussion

The present study is the first to demonstrate that combining methionine restriction with low-dose cisplatinum provides stronger tumor inhibition than standard-dose cisplatinum in both in vitro and in vivo lung-cancer models. Furthermore, in vivo experiments revealed that the combination of a methionine-restricted diet and low-dose cisplatinum caused less body weight loss than standard-dose cisplatinum alone, indicating potentially lower systemic toxicity.

Masaki et al. reported that oral recombinant methioninase (o-rMETase) combined with cisplatinum significantly reduced the effective cisplatinum dose and eliminated cisplatinum-associated toxicity in a patient-derived osteosarcoma nude-mouse model (19). Other studies have reported synergy of rMETase and cisplatinum (15-18).

The present study showed that ultra-low concentrations of cisplatinum, down to the IC10 when combined with rMETase, significantly inhibited A549 lung cancer cells in vitro, an unexpected result. In vivo, low-dose cisplatinum combined with a low-methionine diet eradicated A549 subcutaneous tumors in nude mice.

Methionine addiction, also known as the Hoffman effect, is a fundamental and general hallmark of cancer (7-14). Restricting methionine is known to arrest cancer cells in the S/G2 phase of the cell cycle, which is the same cell-cycle phase targeted by cisplatinum (15, 20). Therefore, the combination of methionine restriction, including rMETase, and cisplatinum has synergistic anti-tumor efficacy in various cancer types.

The absence of substantial weight loss with the combination of low-dose cisplatinum and an MR diet suggests a favorable systemic safety profile and supports the feasibility of dose reduction of cisplatinum when administered in conjunction with methionine restriction. Furthermore, the mechanism of the synergistic efficacy of ultra-low-dose cisplatinum combined with rMETase on cancer cells may be due to extensive chromosomal, nucleolar and nuclear abnormalities in the cancer cells induced by subtoxic concentrations of cisplatinum (24). Further studies on systemic toxicity and organ-specific effects are needed for translation into clinical practice.

Conclusion

The present study provides novel evidence that methionine restriction using rMETase or a low-methionine diet enhances the efficacy of low-dose cisplatinum in lung cancer while maintaining favorable tolerability. The present study suggests ultra-low doses of cisplatinum plus rMETase are effective against lung-cancer cells. This strategy may represent a promising approach to reduce platinum-related toxicity and improve therapeutic outcomes. Further studies are necessary to assess their translational potential in the treatment of lung cancer. rMETase is effective because it targets a fundamental hallmark of cancer (7-14, 20-22, 25-59). rMETase is showing clinical promise (60). Comparison of methionine-based PET imaging with glucose-based PET imaging is showing the Hoffman effect is stronger than the Warburg effect, respectively (61).

Acknowledgements

This article is dedicated to the memory of A.R. Moossa, MD; Professor Philip Miles; Sun Lee, MD; Richard W Erbe, MD; Professor Milton Plesur; Professor Gordon H. Sato; Professor Li Jiaxi; Masaki Kitajima, MD; Joseph R. Bertino, MD; John Mendelsohn, MD; Professor I.J. Fidler; Shigeo Yagi, PhD; J.A.R Mead, PhD. Eugene P. Frenkel, 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

    YA and RMH designed the study. QH and SL provided rMETase. YA performed experiments and formal analysis. YA was the major contributor to writing - original draft and RMH revised the manuscript. QH, SL, BMK, JSK, YM, NY, KH, HK, ShM, KI, TH, SeM, HT, and SD critically read and approved the final manuscript.

  • Conflicts of Interest

    The Authors have no conflicts of interest to declare in relation 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 November 5, 2025.
  • Revision received November 24, 2025.
  • Accepted November 25, 2025.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) 4.0 international license (https://creativecommons.org/licenses/by-nc-nd/4.0).

References

  1. ↵
    1. Bray F,
    2. Laversanne M,
    3. Sung H,
    4. Ferlay J,
    5. Siegel RL,
    6. Soerjomataram I,
    7. Jemal A
    : Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 74(3): 229-263, 2024. DOI: 10.3322/caac.21834
    OpenUrlCrossRefPubMed
  2. ↵
    1. Umar H,
    2. Wahab HA,
    3. Attiq A,
    4. Amjad MW,
    5. Bukhari SNA,
    6. Ahmad W
    : Platinum-based targeted chemotherapies and reversal of cisplatin resistance in non-small cell lung cancer (NSCLC). Mutat Res 828: 111856, 2024. DOI: 10.1016/j.mrfmmm.2024.111856
    OpenUrlCrossRefPubMed
  3. ↵
    1. Chaft JE,
    2. Rimner A,
    3. Weder W,
    4. Azzoli CG,
    5. Kris MG,
    6. Cascone T
    : Evolution of systemic therapy for stages I-III non-metastatic non-small-cell lung cancer. Nat Rev Clin Oncol 18(9): 547-557, 2021. DOI: 10.1038/s41571-021-00501-4
    OpenUrlCrossRef
  4. ↵
    1. Tang C,
    2. Livingston MJ,
    3. Safirstein R,
    4. Dong Z
    : Cisplatin nephrotoxicity: new insights and therapeutic implications. Nat Rev Nephrol 19(1): 53-72, 2023. DOI: 10.1038/s41581-022-00631-7
    OpenUrlCrossRefPubMed
    1. Elmorsy EA,
    2. Saber S,
    3. Hamad RS,
    4. Abdel-Reheim MA,
    5. El-Kott AF,
    6. AlShehri MA,
    7. Morsy K,
    8. Salama SA,
    9. Youssef ME
    : Advances in understanding cisplatin-induced toxicity: Molecular mechanisms and protective strategies. Eur J Pharm Sci 203: 106939, 2024. DOI: 10.1016/j.ejps.2024.106939
    OpenUrlCrossRefPubMed
  6. ↵
    1. McSweeney KR,
    2. Gadanec LK,
    3. Qaradakhi T,
    4. Ali BA,
    5. Zulli A,
    6. Apostolopoulos V
    : Mechanisms of cisplatin-induced acute kidney injury: pathological mechanisms, pharmacological interventions, and genetic mitigations. Cancers (Basel) 13(7): 1572, 2021. DOI: 10.3390/cancers13071572
    OpenUrlCrossRefPubMed
  7. ↵
    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 U S A 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. 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. Kaiser P
    : Methionine dependence of cancer. Biomolecules 10(4): 568, 2020. DOI: 10.3390/biom10040568
    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. Kreis W,
    2. Hession C
    : Biological effects of enzymatic deprivation of L-methionine in cell culture and an experimental tumor. Cancer Res 33(8): 1866-1869, 1973.
    OpenUrlAbstract/FREE Full Text
  14. ↵
    1. Sedillo JC,
    2. Cryns VL
    : Targeting the methionine addiction of cancer. Am J Cancer Res 12(5): 2249-2276, 2022.
    OpenUrlPubMed
  15. ↵
    1. Kubota Y,
    2. Han Q,
    3. Aoki Y,
    4. Masaki N,
    5. Obara K,
    6. Hamada K,
    7. Hozumi C,
    8. Wong ACW,
    9. Bouvet M,
    10. Tsunoda T,
    11. Hoffman RM
    : Synergy of combining methionine restriction and chemotherapy: the disruptive next generation of cancer treatment. Cancer Diagn Progn 3(3): 272-281, 2023. DOI: 10.21873/cdp.10212
    OpenUrlCrossRefPubMed
    1. Hoshiya Y,
    2. Kubota T,
    3. Matsuzaki SW,
    4. Kitajima M,
    5. Hoffman RM
    : Methionine starvation modulates the efficacy of cisplatin on human breast cancer in nude mice. Anticancer Res 16(6B): 3515-3517, 1996.
    OpenUrlPubMed
    1. Asano Y,
    2. Han Q,
    3. Mizuta K,
    4. Kang BM,
    5. Kim JS,
    6. Yamamoto N,
    7. Hayashi K,
    8. Kimura H,
    9. Miwa S,
    10. Igarashi K,
    11. Higuchi T,
    12. Morinaga S,
    13. Tsuchiya H,
    14. Demura S,
    15. Hoffman RM
    : Recombinant methioninase and cisplatinum act synergistically to inhibit lewis lung carcinoma cells but not normal fibroblasts. Anticancer Res 45(5): 1871-1876, 2025. DOI: 10.21873/anticanres.17566
    OpenUrlAbstract/FREE Full Text
  18. ↵
    1. Tan Y,
    2. Sun X,
    3. Xu M,
    4. Tan X,
    5. Sasson A,
    6. Rashidi B,
    7. Han Q,
    8. Tan X,
    9. Wang X,
    10. An Z,
    11. Sun FX,
    12. Hoffman RM
    : Efficacy of recombinant methioninase in combination with cisplatin on human colon tumors in nude mice. Clin Cancer Res 5(8): 2157-2163, 1999.
    OpenUrlAbstract/FREE Full Text
  19. ↵
    1. Masaki N,
    2. Han Q,
    3. Wu NF,
    4. Samonte C,
    5. Wu J,
    6. Hozumi C,
    7. Obara K,
    8. Kubota Y,
    9. Aoki Y,
    10. Miyazaki J,
    11. Hoffman RM
    : Oral-recombinant methioninase lowers the effective dose and eliminates toxicity of cisplatinum for primary osteosarcoma of the mammary gland in a patient-derived orthotopic xenograft mouse model. In Vivo 36(6): 2598-2603, 2022. DOI: 10.21873/invivo.12994
    OpenUrlAbstract/FREE Full Text
  20. ↵
    1. Hoffman RM,
    2. Jacobsen SJ
    : Reversible growth arrest in simian virus 40-transformed human fibroblasts. Proc Natl Acad Sci U S A 77(12): 7306-7310, 1980. DOI: 10.1073/pnas.77.12.7306
    OpenUrlAbstract/FREE Full Text
    1. Wang Y,
    2. Muylaert C,
    3. Wyns A,
    4. Vlummens P,
    5. De Veirman K,
    6. Vanderkerken K,
    7. Zaal E,
    8. Berkers C,
    9. Moreaux J,
    10. De Bruyne E,
    11. Menu E
    : S-adenosylmethionine biosynthesis is a targetable metabolic vulnerability in multiple myeloma. Haematologica 109(1): 256-271, 2024. DOI: 10.3324/haematol.2023.282866
    OpenUrlCrossRefPubMed
  22. ↵
    1. Fujioka S,
    2. Ting RC,
    3. Gallo RC
    : Transfer RNA methylases of normal cells, virus-transformed cells, and tumors derived from transformed cells. Cancer Res 31(4): 451-456, 1971.
    OpenUrlAbstract/FREE Full Text
  23. ↵
    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-mercaptomethanelyase for novel anticancer therapy. Protein Expr Purif 9(2): 233-245, 1997. DOI: 10.1006/prep.1996.0700
    OpenUrlCrossRefPubMed
  24. ↵
    1. Delinassios JG,
    2. Hoffman RM,
    3. Koumakis G,
    4. Palitskaris D,
    5. Poulatsidou KN,
    6. Delinasios GJ
    : Subtoxic cisplatin concentrations induce extensive chromosomal, nuclear and nucleolar abnormalities associated with high malignancy before acquired resistance develops: Implications for clinical caution. PLoS One 19(12): e0311976, 2024. DOI: 10.1371/journal.pone.0311976
    OpenUrlCrossRefPubMed
  25. ↵
    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
    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. 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. 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
    1. Stern PH,
    2. Hoffman RM
    : Elevated overall rates of trans-methylation in cell lines from diverse human tumors. In Vitro 20(8): 663-670, 1984. DOI: 10.1007/bf02619617
    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. 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. 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
    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
    1. Tisdale MJ
    : Utilization of preformed and endogenously synthesized methionine by cells in tissue culture. Br J Cancer 49(3): 315-320, 1984. DOI: 10.1038/bjc.1984.49
    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. Sanderson SM,
    2. Gao X,
    3. Dai Z,
    4. Locasale JW
    : Methionine metabolism in health and cancer: a nexus of diet and precision medicine. Nat Rev Cancer 19(11): 625-637, 2019. DOI: 10.1038/s41568-019-0187-8
    OpenUrlCrossRefPubMed
    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,
    17. Locasale JW
    : Dietary methionine influences therapy in mouse cancer models and alters human metabolism. Nature 572(7769): 397-401, 2019. 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. 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 Commun 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. Kreis W,
    2. Baker A,
    3. Ryan V,
    4. Bertasso A
    : Effect of nutritional and enzymatic methionine deprivation upon human normal and malignant cells in tissue culture. Cancer Res 40(3): 634-641, 1980.
    OpenUrlAbstract/FREE Full Text
    1. Tee T,
    2. Ruiter TJJ,
    3. Wu S,
    4. Zhang W,
    5. Schenau DVI,
    6. Rodionova M,
    7. Wajon D,
    8. Vervoort BMT,
    9. Grunewald KJT,
    10. Bosma M,
    11. Hagelaar R,
    12. Baker-Hernandez J,
    13. Dahaoui A,
    14. Schneider P,
    15. Verhoeven-Duif NM,
    16. Van der Meer LT,
    17. Van Leeuwen FN
    : S-adenosylmethionine addiction confers sensitivity to methionine restriction in KMT2A-rearranged acute lymphoblastic leukemia. Haematologica 110(11): 2620-2634, 2025. DOI: 10.3324/haematol.2023.284869
    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. Kreis W
    : Tumor therapy by deprivation of L-methionine: rationale and results. Cancer Treat Rep. 63(6): 1069-1072, 1979.
    OpenUrlPubMed
    1. Breillout F,
    2. Hadida F,
    3. Echinard-Garin P,
    4. Lascaux V,
    5. Poupon MF
    : Decreased rat rhabdomyosarcoma pulmonary metastases in response to a low methionine diet. Anticancer Res 7(4B): 861-867, 1987.
    OpenUrlPubMed
    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. Poirson-Bichat F,
    2. Lopez R,
    3. Bras Gonçalves RA,
    4. Miccoli L,
    5. Bourgeois Y,
    6. Demerseman P,
    7. Poisson M,
    8. Dutrillaux B,
    9. Poupon MF
    : Methionine deprivation and methionine analogs inhibit cell proliferation and growth of human xenografted gliomas. Life Sci 60(12): 919-931, 1977. DOI: 10.1016/s0024-3205(96)00672-8
    OpenUrlCrossRef
    1. Poirson-Bichat F,
    2. Gonfalone G,
    3. Bras-Gonçalves RA,
    4. Dutrillaux B,
    5. Poupon MF
    : Growth of methionine-dependent human prostate cancer (PC-3) is inhibited by ethionine combined with methionine starvation. Br J Cancer 75(11): 1605-1612, 1977. DOI: 10.1038/bjc.1997.274
    OpenUrlCrossRef
    1. Poirson-Bichat F,
    2. Gonçalves RA,
    3. Miccoli L,
    4. Dutrillaux B,
    5. Poupon MF
    : Methionine depletion enhances the antitumoral efficacy of cytotoxic agents in drug-resistant human tumor xenografts. Clin Cancer Res 6(2): 643-653, 2000.
    OpenUrlAbstract/FREE Full Text
    1. Lauinger L,
    2. Kaiser P
    : Sensing and signaling of methionine metabolism. Metabolites 11(2): 83, 2021. DOI: 10.3390/metabo11020083
    OpenUrlCrossRefPubMed
    1. Borrego SL,
    2. Lin DW,
    3. Kaiser P
    : Isolation and characterization of methionine-independent clones from methionine-dependent cancer cells. Methods Mol Biol 1866: 37-48, 2019. DOI: 10.1007/978-1-4939-8796-2_4
    OpenUrlCrossRef
    1. Malin D,
    2. Lee Y,
    3. Chepikova O,
    4. Strekalova E,
    5. Carlson A,
    6. Cryns VL
    : Methionine restriction exposes a targetable redox vulnerability of triple-negative breast cancer cells by inducing thioredoxin reductase. Breast Cancer Res Treat 190(3): 373-387, 2021. DOI: 10.1007/s10549-021-06398-y
    OpenUrlCrossRefPubMed
    1. Haws SA,
    2. Yu D,
    3. Ye C,
    4. Wille CK,
    5. Nguyen LC,
    6. Krautkramer KA,
    7. Tomasiewicz JL,
    8. Yang SE,
    9. Miller BR,
    10. Liu WH,
    11. Igarashi K,
    12. Sridharan R,
    13. Tu BP,
    14. Cryns VL,
    15. Lamming DW,
    16. Denu JM
    : Methyl-metabolite depletion elicits adaptive responses to support heterochromatin stability and epigenetic persistence. Mol Cell 78(2): 210-223.e8, 2020. DOI: 10.1016/j.molcel.2020.03.004
    OpenUrlCrossRefPubMed
  59. ↵
    1. Rajanala SH,
    2. Ringquist R,
    3. Cryns VL
    : Methionine restriction activates the integrated stress response in triple-negative breast cancer cells by a GCN2- and PERK-independent mechanism. Am J Cancer Res 9(8): 1766-1775, 2019.
    OpenUrlPubMed
  60. ↵
    1. Asano Y,
    2. Han Q,
    3. Li S,
    4. Mizuta K,
    5. Kang BM,
    6. Kim JS,
    7. Miyashi Y,
    8. Yamamoto N,
    9. Hayashi K,
    10. Kimura H,
    11. Miwa S,
    12. Igarashi K,
    13. Higuchi T,
    14. Morinaga S,
    15. Tsuchiya H,
    16. Demura S,
    17. Hoffman RM
    : Very rapid eradication of a large squamous-cell carcinoma of the head and neck treated with first-line combination chemotherapy, a low-methionine diet, and oral recombinant methioninase. Anticancer Res 45(11): 5225-5231, 2025. DOI: 10.21873/anticanres.17862
    OpenUrlAbstract/FREE Full Text
  61. ↵
    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 wholebody imaging to determine the extent of methionineaddiction 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
Back to top

In this issue

Print
Download PDF
Article Alerts
Email Article
Citation Tools
Reprints and Permissions
Ultra-low Concentrations of Cisplatinum Down to the IC10 in Combination With Recombinant Methioninase Are Synergistically Effective Against Lung Cancer Cells In Vitro and In Vivo
YOHEI ASANO, QINGHONG HAN, SHUKUAN LI, BYUNG MO KANG, JIN SOO KIM, YUTA MIYASHI, MICHAEL BOUVET, NORIO YAMAMOTO, KATSUHIRO HAYASHI, HIROAKI KIMURA, SHINJI MIWA, KENTARO IGARASHI, TAKASHI HIGUCHI, SEI MORINAGA, HIROYUKI TSUCHIYA, SATORU DEMURA, ROBERT M. HOFFMAN
In Vivo Mar 2026, 40 (2) 803-812; DOI: 10.21873/invivo.14238
Twitter logo Facebook logo Mendeley logo

Jump to section

Keywords