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Targeting of a novel fusion protein containing methioninase to the urokinase receptor to inhibit breast cancer cell migration and proliferation

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Abstract

It has been shown that methionine depletion inhibits tumor cell growth and reduces tumor cell survival. A novel fusion protein targeted specifically to tumor cells was developed. The fusion protein contained two components: the amino terminal fragment of human urokinase (amino acids 1–49) that binds to the urokinase receptor protein expressed on the surface of invasive cancer cells, and the enzyme l-methioninase (containing 398 amino acids) which depletes methionine and arrests the growth of methionine-dependent tumors. The influence of the fusion protein on the growth and motility of human breast cancer cells was examined using a culture wounding assay. It was determined that MCF-7 breast cancer cells, used in this study, were methionine-dependent and that the fusion protein bound specifically to urokinase receptors of the surface of the cancer cells. Further treatment of the cancer cells with fusion protein over the concentration range 10−8 to 10−6  M produced a dose-dependent inhibition of both the migration and proliferation index of MCF-7 cells in the culture wounding assay over a period of 1 to 3 days. The results of this study suggest that this novel fusion protein may serve as a prototype for specific targeting of methioninase and perhaps other cytotoxic agents to cancer cells.

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Abbreviations

ATF:

Amino-terminal fragment of urokinase (amino acids 1–49)

PMSF:

Phenylmethylsulfonyl fluoride

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

TPCK:

N-p-Tosyl-l-phenylalanine chloromethyl ketone

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Acknowledgements

The authors acknowledge the support of the University of Oklahoma, Bioengineering Center. This work was also supported in part by grants from NIH/NCI (CA-89740), DoD (DAMD17-01-1-0591) and OCAST (HR03-046).

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Authors and Affiliations

  1. Bioengineering Center and the School of Chemical Engineering and Materials Science, University of Oklahoma, Norman, Oklahoma, 73019, USA

    Karine Peron, Sebastien A. Gauthier, Magali Barriere, Damien Prévéraud, Charles E. Soliman & Roger G. Harrison

  2. Department of Pharmaceutical Science, College of Pharmacy, Health Sciences Center, University of Oklahoma, Oklahoma City, Oklahoma, 73117, USA

    Tara N. Jones, Thao-Nguyen T. Nguyen, Xiao-Ping Zang & J. Thomas Pento

Authors
  1. Karine Peron
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  2. Tara N. Jones
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  3. Sebastien A. Gauthier
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  4. Thao-Nguyen T. Nguyen
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  5. Xiao-Ping Zang
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  6. Magali Barriere
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  7. Damien Prévéraud
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  8. Charles E. Soliman
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  9. Roger G. Harrison
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  10. J. Thomas Pento
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Corresponding author

Correspondence to J. Thomas Pento.

Additional information

K.P. and S.A.G. are intern students from Blaise Pascal University, Clermont-Ferrand, France. M.B. and D.P. are intern students from the University of Limoges, France.

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Peron, K., Jones, T.N., Gauthier, S.A. et al. Targeting of a novel fusion protein containing methioninase to the urokinase receptor to inhibit breast cancer cell migration and proliferation. Cancer Chemother Pharmacol 52, 270–276 (2003). https://doi.org/10.1007/s00280-003-0666-0

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  • DOI: https://doi.org/10.1007/s00280-003-0666-0

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