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. 2025 Jan-Feb;39(1):120-126.
doi: 10.21873/invivo.13809.

Synergistic Eradication of Fibrosarcoma With Acquired Ifosfamide Resistance Using Methionine Restriction Combined With Ifosfamide in Nude-mouse Models

Sei Morinaga  1   2   3 , Qinghong Han  1 , Kohei Mizuta  1   2 , Byung Mo Kang  1   2 , Michael Bouvet  2 , Norio Yamamoto  3 , Katsuhiro Hayashi  3 , Hiroaki Kimura  3 , Shinji Miwa  3 , Kentaro Igarashi  3 , Takashi Higuchi  3 , Hiroyuki Tsuchiya  3 , Satoru Demura  3 , Robert M Hoffman  4   2
Affiliations

Synergistic Eradication of Fibrosarcoma With Acquired Ifosfamide Resistance Using Methionine Restriction Combined With Ifosfamide in Nude-mouse Models

Sei Morinaga et al. In Vivo. 2025 Jan-Feb.

Abstract

Background/aim: Ifosfamide is used clinically with doxorubicin as first-line chemotherapy for soft-tissue sarcoma. However, ifosfamide efficacy for soft-tissue sarcoma is limited due to frequent occurence of ifosfamide resistance and thus more effective therapy is needed. The present study aimed to determine the synergy of recombinant methioninase (rMETase) plus ifosfamide against HT1080 human fibrosarcoma cells in vitro. Additionally, the present study also investigated the efficacy of a methionine-restricted diet combined with ifosfamide in nude-mouse models of ifosfamide-resistant HT1080 (IR-HT1080).

Materials and methods: Cell viability for HT1080 human fibrosarcoma cells was determined in four groups in vitro: No treatment control; ifosfamide alone; rMETase alone; and a combination of ifosfamide plus rMETase. HT1080 tumors were established in nude mice subcutaneously. The HT1080 tumor models were treated by administering ifosfamide by intraperitoneal injection twice a week, for a total of 11 doses. Surviving tumors were considered ifosfamide resistant (IR-HT1080). Four groups of IR-HT1080 nude-mouse models were subsequently established: Group 1 was a no-treatment control, Group 2 received ifosfamide, Group 3 was given a methionine-restricted diet (MR), and Group 4 received ifosfamide plus MR. Additionally, two groups of nude mice with parental HT1080 subcutaneous tumors were included: Group 5 was a no-treatment control, and Group 6 received ifosfamide for comparison.

Results: The 50% inhibitory concentration (IC50) for ifosfamide against HT1080 cells was 0.38 mM. The IC50 for rMETase was 0.75 U/ml for HT1080 cells (data from [4]). The combination of rMETase (0.75 U/ml) plus ifosfamide (0.38 mM) was synergistic against HT1080 fibrosarcoma cells in vitro. The combination of ifosfamide plus MR eradicated the IR-HT1080 tumors in nude-mouse models, while each treatment alone achieved limited tumor inhibition.

Conclusion: The present results suggest the combination of MR and ifosfamide has promising potential for overcoming ifosfamide resistance in future clinical applications.

Keywords: HT1080; Hoffman effect; Methioninase; fibrosarcoma; ifosfamide; ifosfamide-resistance; methionine restriction; synergy; tumor eradication methionine addiction.

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Conflict of interest statement

The Authors declare no competing interests in relation to this study.

Figures

Figure 1
Figure 1
Ifosfamide and rMETase sensitivity of HT1080 fibrosarcoma cells in vitro (mean±standard deviation). A) Sensitivity of HT1080 cells to ifosfamide. B) Sensitivity of HT1080 cells to rMETase.
Figure 2
Figure 2
Efficacy of the combination of ifosfamide and rMETase, alone and in combination on HT1080 fibrosarcoma cells. Control (DMEM); ifosfamide [0.38 mM (IC50)]; rMETase [0.75 U/ml (IC50)]; ifosfamide [0.38 mM (IC50)] and rMETase [0.75 U/ml (IC50)].
Figure 3
Figure 3
The combination of ifosfamide plus methionine restriction eradicated ifosfamide-resistant HT1080 fibrosarcoma (IR-HT1080) in nude mice. A) Time course efficacy of methionine-restricted diet (MR) and ifosfamied alone and in combination on ifosfamide-resistant (IR)-HT1080 tumors in nude mice. Ifosfamide was also tested on HT1080 tumors compared to a no-treatment control. B) IR-HT1080 and HT1080 tumor volume on day 14 for each treatment group. Data are shown as the mean±standard deviation. Please see Materials and Methods for dose, route and schedule of ifosfamide treatment.
Figure 4
Figure 4
Effect of the ifosfamide and a methionine-restricted (MR) diet alone and in combination on the body weight of nude mice with ifosfamide-resistant HT1080 (IR-HT1080) and parental HT1080 tumors. Data are shown as the mean±standard deviation. Untreated control (IR-HT1080); ifosfamide (IR-HT1080); methionine-restricted diet (MR) (IR-HT1080); ifosfamide plus methionine restriction (IR-HT1080); untreated control (HT1080); ifosfamide (HT1080).
See this image and copyright information in PMC

References

    1. Judson I, Verweij J, Gelderblom H, Hartmann JT, Schöffski P, Blay JY, Kerst JM, Sufliarsky J, Whelan J, Hohenberger P, Krarup-Hansen A, Alcindor T, Marreaud S, Litière S, Hermans C, Fisher C, Hogendoorn PC, dei Tos AP, van der Graaf WT, European Organisation and Treatment of Cancer Soft Tissue and Bone Sarcoma Group Doxorubicin alone versus intensified doxorubicin plus ifosfamide for first-line treatment of advanced or metastatic soft-tissue sarcoma: a randomised controlled phase 3 trial. Lancet Oncol. 2014;15(4):415–423. doi: 10.1016/S1470-2045(14)70063-4. - DOI - PubMed
    1. Tan Y, Xu M, Tan X, Tan X, Wang X, Saikawa Y, Nagahama T, Sun X, Lenz M, Hoffman RM. Overexpression and large-scale production of recombinantl-methionine-α-deamino-γ-mercaptomethane-lyase for novel anticancer therapy. Protein Expr Purif. 1997;9(2):233–245. doi: 10.1006/prep.1996.0700. - DOI - PubMed
    1. Choobin BB, Kubota Y, Han Q, Ardjmand D, Morinaga S, Mizuta K, Bouvet M, Tsunoda T, Hoffman RM. Recombinant methioninase lowers the effective dose of regorafenib against colon-cancer cells: a strategy for widespread clinical use of a toxic drug. Cancer Diagn Progn. 2023;3(6):655–659. doi: 10.21873/cdp.10268. - DOI - PMC - PubMed
    1. Morinaga S, Han Q, Kubota Y, Mizuta K, Kang BM, Sato M, Bouvet M, Yamamoto N, Hayashi K, Kimura H, Miwa S, Igarashi K, Higuchi T, Tsuchiya H, Hoffman RM. Extensive synergy between recombinant methioninase and eribulin against fibrosarcoma cells but not normal fibroblasts. Anticancer Res. 2024;44(3):921–928. doi: 10.21873/anticanres.16886. - DOI - PubMed
    1. Ardjmand D, Kubota Y, Sato M, Han Q, Mizuta K, Morinaga S, Hoffman RM. Selective synergy of rapamycin combined with methioninase on cancer cells compared to normal cells. Anticancer Res. 2024;44(3):929–933. doi: 10.21873/anticanres.16887. - DOI - PubMed

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