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Information Journal Paper

Title

CYTOTOXIC EFFECT OF IMMUNOTOXIN CONTAINING THE TRUNCATED FORM OF PSEUDOMONAS EXOTOXIN A AND ANTI-VEGFR2 ON HUVEC AND MCF-7 CELL LINES

Pages

  203-210

Abstract

 Objective: IMMUNOTOXINs (ITs) have been developed for the treatment of cancer, and comprise of antibodies linked to toxins. Also vascular endothelial growth factor (VEGF) plays a key role in tumor angiogenesis, and the blockade of VEGF receptor-2 (VEGFR2) inhibits angiogenesis and tumor growth. The aim of this study was to produce anti-VEGFR2/rPE (Pseudomonas exotoxin) 38 IT to test its cytotoxic activity and mechanism of action.Materials and Methods: In this basic research and experimental study, at first, DNA that encodes recombinant PE38 protein was inductively expressed in Escherichia coli (E.coli) and purified by nickel-sepharose chromatography and further analyzed by western blot. Then, for production of IT, rPE38 was chemically conjugated to anti-VEGFR2. The cytotoxicity response of IT treatment was evaluated by 3- (4, 5-Dimethylthiazol-2-Yl) -2, 5-Diphenyltetrazolium Bromide (MTT) test in Human Umbilical Vein Endothelial Cell (HUVEC) and Michigan Cancer Foundation-7 (MCF-7) (VEGFR2+) cell lines. The mechanism of IT cytotoxicity was observed by Annexin V staining and flow cytometry. Continuous variables were compared with the analysis of variance (ANOVA; for all groups). P values less than 0.05 were considered statistically significant.Results: SDS-PAGE showed 98% purity of rPE38 and IT. In vitro dose-dependent cytotoxicity assay demonstrated that anti-VEGFR2/PE38 is toxic to VEGFR2-positive cells. IT treatment significantly inhibited proliferation of HUVEC and MCF-7 in a VEGFR2-specific manner as compared with the control groups (p<0.05). Flow cytometry showed that the mechanism of IT induced cell death is mediated by apoptosis.Conclusion: IT treatment also caused remarkable synergistic cytotoxicity characterized by decreased cell viability, and an increased apoptotic index by both anti-VEGFR2 and PE38. Thus these results raise the possibility of using anti-VEGFR2/PE38 IT for cancer therapy because nearly all tumors induce local angiogenesis with high VEGFR expression.

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    APA: Copy

    SAFARI, ELAHE, ZAVARAN HOSSEINI, AHMAD, HASSAN, ZUHAIR, KHAJEH, KHOSRO, SHAFIEE ARDESTANI, MEHDI, & BARADARAN, BEHZAD. (2014). CYTOTOXIC EFFECT OF IMMUNOTOXIN CONTAINING THE TRUNCATED FORM OF PSEUDOMONAS EXOTOXIN A AND ANTI-VEGFR2 ON HUVEC AND MCF-7 CELL LINES. CELL JOURNAL (YAKHTEH), 16(2 (62)), 203-210. SID. https://sid.ir/paper/660232/en

    Vancouver: Copy

    SAFARI ELAHE, ZAVARAN HOSSEINI AHMAD, HASSAN ZUHAIR, KHAJEH KHOSRO, SHAFIEE ARDESTANI MEHDI, BARADARAN BEHZAD. CYTOTOXIC EFFECT OF IMMUNOTOXIN CONTAINING THE TRUNCATED FORM OF PSEUDOMONAS EXOTOXIN A AND ANTI-VEGFR2 ON HUVEC AND MCF-7 CELL LINES. CELL JOURNAL (YAKHTEH)[Internet]. 2014;16(2 (62)):203-210. Available from: https://sid.ir/paper/660232/en

    IEEE: Copy

    ELAHE SAFARI, AHMAD ZAVARAN HOSSEINI, ZUHAIR HASSAN, KHOSRO KHAJEH, MEHDI SHAFIEE ARDESTANI, and BEHZAD BARADARAN, “CYTOTOXIC EFFECT OF IMMUNOTOXIN CONTAINING THE TRUNCATED FORM OF PSEUDOMONAS EXOTOXIN A AND ANTI-VEGFR2 ON HUVEC AND MCF-7 CELL LINES,” CELL JOURNAL (YAKHTEH), vol. 16, no. 2 (62), pp. 203–210, 2014, [Online]. Available: https://sid.ir/paper/660232/en

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