title: Antiproliferative and apoptotic effects of a specific anti-insulin-like growth factor I receptor single chain antibody on breast cancer cells.
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 VHand 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.
authors.affiliation: Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.|Department of Immunology, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Morteza
authors.initials: M
authors.lastname: Motallebnezhad
authors.affiliation: Faculty of Paramedical Sciences, Alborz University of Medical Sciences, Karaj, Iran. v.younesi@pishtazteb.com.|Pishtaz Teb Diagnostics, Tehran, Iran. v.younesi@pishtazteb.com.
authors.forename: Vahid
authors.initials: V
authors.lastname: Younesi
authors.affiliation: Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.|Department of Immunology, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Leili
authors.initials: L
authors.lastname: Aghebati-Maleki
authors.affiliation: Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.|Department of Immunology, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Hamid
authors.initials: H
authors.lastname: Nickho
authors.affiliation: Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.|Department of Immunology, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Elham
authors.initials: E
authors.lastname: Safarzadeh
authors.affiliation: Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.|Department of Immunology, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Ahmadi
authors.affiliation: Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Ali Akbar
authors.initials: AA
authors.lastname: Movassaghpour
authors.affiliation: Shiraz Institute for Cancer Research, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Hosseini
authors.affiliation: Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. yousefime@tbzmed.ac.ir.|Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran. yousefime@tbzmed.ac.ir.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Yousefi
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000188:drug therapy*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D006801:Humans; D061986:MCF-7 Cells; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000037:antagonists & inhibitors*; D017526:Receptor, IGF Type 1 / Q000037:antagonists & inhibitors* / Q000276:immunology; D057127:Single-Chain Antibodies / Q000276:immunology / Q000494:pharmacology*; D000068878:Trastuzumab / Q000494:pharmacology*
chemical_list: D000911:Antibodies, Monoclonal; D018719:Erb-b2 Receptor Tyrosine Kinases; D017526:Receptor, IGF Type 1; D057127:Single-Chain Antibodies; D000068878:Trastuzumab; C508053:ERBB2 protein, human
keywords: Breast cancer; Insulin-like growth factor I receptor; Phage display; Single-chain fragment variable (scFv) antibodies; Trastuzumab