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Tumor Biology

, Volume 37, Issue 11, pp 14841–14850 | Cite as

Antiproliferative and apoptotic effects of a specific anti-insulin-like growth factor I receptor single chain antibody on breast cancer cells

  • Morteza Motallebnezhad
  • Vahid Younesi
  • Leili Aghebati-Maleki
  • Hamid Nickho
  • Elham Safarzadeh
  • Majid Ahmadi
  • Ali Akbar Movassaghpour
  • Ahmad Hosseini
  • Mehdi Yousefi
Original Article

Abstract

Insulin-like growth factor I receptor (IGF-IR) is expressed on breast cancer cells and involves in metastasis, survival, and proliferation. Currently, application of IGF-IR-targeting monoclonal antibodies (mAbs), alone or in combination with other drugs, is a promising strategy for breast cancer therapy. Single-chain fragment variable (scFv) antibodies have been introduced as appropriate tools for tumor-targeting purposes because of their advantages over whole antibodies. In the present study, we employed a naïve phage library and isolated scFvs against a specific epitope from extracellular domain of IGF-IR by panning process. The selected scFvs were further characterized using polyclonal and monoclonal phage ELISA, soluble monoclonal ELISA, and colony PCR and sequencing. Antiproliferative and apoptotic effects of selected scFv antibodies on breast cancer cell lines were also evaluated by MTT and Annexin V/PI assays. The results of ELISA indicated specific reactions of the isolated scFvs against the IGF-IR peptide, and analyses of PCR product and sequencing confirmed the presence of full length VH and Vκ inserts. Treatment of MCF7 and SKBR3 cells with anti-IGF-IR scFv led to a significant growth inhibition. The results also showed that scFv treatment significantly augmented trastuzumab growth inhibitory effects on SKBR3 cells. The percentage of the apoptotic MCF7 and SKBR3 cells after 24-h treatment with scFv was 39 and 30.70 %, respectively. Twenty-four-hour treatment with scFv in combination with trastuzumab resulted in 44.75 % apoptosis of SKBR3 cells. Taken together, our results demonstrate that the targeting of IGF-IR by scFv can be an effective strategy in the treatment of breast cancer and provide further evidence for effectiveness of dual targeting of HER2 and IGF-IR in breast cancer therapy.

Keywords

Insulin-like growth factor I receptor Phage display Single-chain fragment variable (scFv) antibodies Breast cancer Trastuzumab 

Notes

Acknowledgments

This work was financially supported by the Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Compliance with ethical standards

Conflicts of interest

None

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Morteza Motallebnezhad
    • 1
    • 2
  • Vahid Younesi
    • 3
    • 4
  • Leili Aghebati-Maleki
    • 1
    • 2
  • Hamid Nickho
    • 1
    • 2
  • Elham Safarzadeh
    • 1
    • 2
  • Majid Ahmadi
    • 1
    • 2
  • Ali Akbar Movassaghpour
    • 5
  • Ahmad Hosseini
    • 6
  • Mehdi Yousefi
    • 7
    • 8
  1. 1.Immunology Research CenterTabriz University of Medical SciencesTabrizIran
  2. 2.Department of Immunology, School of MedicineTabriz University of Medical SciencesTabrizIran
  3. 3.Faculty of Paramedical SciencesAlborz University of Medical SciencesKarajIran
  4. 4.Pishtaz Teb DiagnosticsTehranIran
  5. 5.Hematology and Oncology Research CenterTabriz University of Medical SciencesTabrizIran
  6. 6.Shiraz Institute for Cancer ResearchShiraz University of Medical SciencesShirazIran
  7. 7.Drug Applied Research CenterTabriz University of Medical SciencesTabrizIran
  8. 8.Department of Immunology, Faculty of MedicineTabriz University of Medical SciencesTabrizIran

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