Skip to main content

The inhibition of glutamine synthetase sensitizes human sarcoma cells to l-asparaginase

Cancer Chemotherapy and Pharmacology volume 60, pages 751–758 (2007)Cite this article

Abstract

Purpose

To evaluate the activity of the antitumor enzyme l-asparaginase (ASNase) on tumor cells of mesenchymal origin and the contribution of glutamine synthetase (GS) to the adaptation to the metabolic stress caused by the anti-tumor enzyme.

Methods

We studied the effects of ASNase in six human sarcoma cell lines: HT1080 (fibrosarcoma); RD (rhabdomyosarcoma); SW872 (liposarcoma); HOS, SAOS-2, and U2OS (osteosarcoma) in the absence or in the presence of the GS inhibitor methionine l-sulfoximine (MSO).

Results

HT1080 and SW872 cells were highly sensitive to ASNase-dependent cytotoxicity. In contrast, RD, SAOS-2, HOS, and U2OS cells exhibited only a partial growth suppression upon treatment with the anti-tumor enzyme. In these cell lines ASNase treatment was associated with increased levels of GS. When ASNase was used together with MSO, the proliferation of the poorly sensitive cell lines was completely blocked and a significant decrease in the IC50 for ASNase was observed. Moreover, when ASNase treatment was carried on in the presence of MSO, HOS and U2OS osteosarcoma cells exhibited a marked cytotoxicity, with increased apoptosis.

Conclusions

In human sarcoma cells (1) GS markedly contributes to the metabolic adaptation of tumor cells to ASNase and (2) the inhibition of GS activity enhances the antiproliferative and cytotoxic effects of ASNase. The two-step interference with glutamine metabolism, obtained through the combined treatment with ASNase and MSO, may provide a novel therapeutic approach that should be further investigated in human tumors of mesenchymal origin.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Abbreviations

ASNase :

l-Asparaginase

AS :

Asparagine synthetase

DMEM :

Dulbecco’s modified Eagle’s medium

FBS :

Fetal bovine serum

GS :

Glutamine synthetase

MSO :

Methionine-l-sulfoximine

References

  1. Aghaiypour K, Wlodawer A, Lubkowski J (2001) Structural basis for the activity and substrate specificity of Erwinia chrysanthemi l-asparaginase. Biochemistry 40:5655–5664

    PubMed  Article  CAS  Google Scholar 

  2. Ando M, Sugimoto K, Kitoh T et al (2005) Selective apoptosis of natural killer-cell tumours by l-asparaginase. Br J Haematol 130:860–868

    PubMed  Article  CAS  Google Scholar 

  3. Aslanian AM, Kilberg MS (2001) Multiple adaptive mechanisms affect asparagine synthetase substrate availability in asparaginase-resistant MOLT-4 human leukaemia cells. Biochem J 358:59–67

    PubMed  Article  CAS  Google Scholar 

  4. Asselin BL, Ryan D, Frantz CN et al (1989) In vitro and in vivo killing of acute lymphoblastic leukemia cells by l-asparaginase. Cancer Res 49:4363–4368

    PubMed  CAS  Google Scholar 

  5. Bussolati O, Belletti S, Uggeri J et al (1995) Characterization of apoptotic phenomena induced by treatment with l-asparaginase in NIH3T3 cells. Exp Cell Res 220:283–291

    PubMed  Article  CAS  Google Scholar 

  6. Fine BM, Kaspers GJ, Ho M, Loonen AH, Boxer LM (2005) A genome-wide view of the in vitro response to l-asparaginase in acute lymphoblastic leukemia. Cancer Res 65:291–299

    PubMed  CAS  Google Scholar 

  7. Holleman A, Cheok MH, den Boer ML et al (2004) Gene-expression patterns in drug-resistant acute lymphoblastic leukemia cells and response to treatment. N Engl J Med 351:533–542

    PubMed  Article  CAS  Google Scholar 

  8. Jagoe RT, Lecker SH, Gomes M, Goldberg AL (2002) Patterns of gene expression in atrophying skeletal muscles: response to food deprivation. Faseb J 16:1697–1712

    PubMed  Article  CAS  Google Scholar 

  9. Krejci O, Starkova J, Otova B et al (2004) Upregulation of asparagine synthetase fails to avert cell cycle arrest induced by l-asparaginase in TEL/AML1-positive leukaemic cells. Leukemia 18:434–441

    PubMed  Article  CAS  Google Scholar 

  10. Kufe DW, Pollock RE, Weichselbaum RR, Bast RC Jr, Gansler TS, Holland JF, Frei E III (2003) Holland-Frei Cancer Medicine. B.C.Decker, Hamilton, Ontario, cap. 55

  11. Labow BI, Souba WW, Abcouwer SF (2001) Mechanisms governing the expression of the enzymes of glutamine metabolism–glutaminase and glutamine synthetase. J Nutr 131:2467S–2474S; discussion 2486S–2467S

    Google Scholar 

  12. Lacoste L, Chaudhary KD, Lapointe J (1982) Derepression of the glutamine synthetase in neuroblastoma cells at low concentrations of glutamine. J Neurochem 39:78–85

    PubMed  Article  CAS  Google Scholar 

  13. Lazarus H, McCoy TA, Farber S, Barell EF, Foley GE (1969) Nutritional requirements of human leukemic cells. Asparagine requirements and the effect of l-asparaginase. Exp Cell Res 57:134–138

    PubMed  Article  CAS  Google Scholar 

  14. Minet R, Villie F, Marcollet M, Meynial-Denis D, Cynober L (1997) Measurement of glutamine synthetase activity in rat muscle by a colorimetric assay. Clin Chim Acta 268:121–132

    PubMed  Article  CAS  Google Scholar 

  15. Muller HJ, Boos J (1998) Use of l-asparaginase in childhood ALL. Crit Rev Oncol Hematol 28:97–113

    PubMed  Article  CAS  Google Scholar 

  16. O’Brien J, Wilson I, Orton T, Pognan F (2000) Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem 267:5421–5426

    PubMed  Article  CAS  Google Scholar 

  17. Okada S, Hongo T, Yamada S et al (2003) In vitro efficacy of l-asparaginase in childhood acute myeloid leukaemia. Br J Haematol 123:802–809

    PubMed  Article  CAS  Google Scholar 

  18. Olkku A, Bodine PV, Linnala-Kankkunen A, Mahonen A (2004) Glucocorticoids induce glutamine synthetase expression in human osteoblastic cells: a novel observation in bone. Bone 34:320–329

    PubMed  Article  CAS  Google Scholar 

  19. Ollenschlager G, Roth E, Linkesch W et al (1988) Asparaginase-induced derangements of glutamine metabolism: the pathogenetic basis for some drug-related side-effects. Eur J Clin Invest 18:512–516

    PubMed  CAS  Article  Google Scholar 

  20. Reinert RB, Oberle LM, Wek SA et al (2006) Role of glutamine depletion in directing tissue-specific nutrient stress responses to l-asparaginase. J Biol Chem 281(42):31222–31233

    PubMed  Article  CAS  Google Scholar 

  21. Rotoli BM, Uggeri J, Dall’Asta V et al (2005) Inhibition of glutamine synthetase triggers apoptosis in asparaginase-resistant cells. Cell Physiol Biochem 15:281–292

    PubMed  Article  CAS  Google Scholar 

  22. Rudman D, Vogler WR, Howard CH, Gerron GG (1971) Observations on the plasma amino acids of patients with acute leukemia. Cancer Res 31:1159–1165

    PubMed  CAS  Google Scholar 

  23. Stams WA, den Boer ML, Beverloo HB et al (2003) Sensitivity to l-asparaginase is not associated with expression levels of asparagine synthetase in t(12;21) + pediatric ALL. Blood 101:2743–2747

    PubMed  Article  CAS  Google Scholar 

  24. Tezuka K, Nakayama H, Honda K et al (2002) Treatment of a child with myeloid/NK cell precursor acute leukemia with l-asparaginase and unrelated cord blood transplantation. Int J Hematol 75:201–206

    PubMed  Article  Google Scholar 

  25. Uggeri J, Belletti S, Bussolati O, Dall’Asta V, Gazzola GC (1995) Suppression of anionic amino acid transport impairs the maintenance of intracellular glutamate in Ha-ras-expressing cells. Biochem Biophys Res Commun 211:878–884

    PubMed  Article  CAS  Google Scholar 

  26. Viceps-Madore D, Cristofalo VJ (1978) Age-associated changes in glutamine synthetase activity in WI-38 cells. Mech Ageing Dev 8:43–50

    PubMed  Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by FIL—University of Parma.

Author information

Affiliations

  1. Unit of General and Clinical Pathology, Department of Experimental Medicine, University of Parma, Via Volturno 39, 43100, Parma, Italy

    S. Tardito, M. G. Bianchi, B. M. Rotoli, R. Franchi-Gazzola, G. C. Gazzola & O. Bussolati

  2. Unit of Histology, Department of Experimental Medicine, University of Parma, Via Volturno 39, 43100, Parma, Italy

    J. Uggeri & R. Gatti

  3. Department of Medical Oncology, University Hospital, Via Gramsci 14, 43100, Parma, Italy

    C. Bozzetti

Authors
  1. S. Tardito
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Uggeri
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Bozzetti
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. G. Bianchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. B. M. Rotoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. R. Franchi-Gazzola
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. G. C. Gazzola
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. R. Gatti
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. O. Bussolati
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to O. Bussolati.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Tardito, S., Uggeri, J., Bozzetti, C. et al. The inhibition of glutamine synthetase sensitizes human sarcoma cells to l-asparaginase. Cancer Chemother Pharmacol 60, 751–758 (2007). https://doi.org/10.1007/s00280-007-0421-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00280-007-0421-z

Keywords

  • Asparaginase
  • Glutamine synthetase
  • HT1080
  • Methionine sulfoximine
  • Osteosarcoma
  • Rhabdomyosarcoma