Evaluation of Cytotoxic and Apoptotic Effects of DT386–BR2: A Promising Anticancer Fusion Protein

authors:

avatar Fatemeh Shafiee 1 , avatar Mohammad Rabbani Khorasgani 1 , avatar Ali Jahanian-Najafabadi 1 , *

Department of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran

how to cite: Shafiee F, Rabbani Khorasgani M, Jahanian-Najafabadi A. Evaluation of Cytotoxic and Apoptotic Effects of DT386–BR2: A Promising Anticancer Fusion Protein. J Rep Pharm Sci. 2020;9(1):e147284. https://doi.org/10.4103/jrptps.JRPTPS_15_19.

Abstract

Purpose: In the previous studies, we designed an anticancer immunotoxin containing the catalytic and translocation domains of diphtheria toxin fused to BR2, a buforin II-derived antimicrobial peptide as a cancer-specific cell penetrating peptide, in order to target various cancer cells. The aim of this study was to evaluate the in vitro cytotoxicity of DT386–BR2 against K-562 cells as the most famous cell line for leukemia. 
Materials and Methods: MTT and flow-cytometry assays were used for determining the cytotoxic effects and cell death mechanism of DT386–BR2, respectively, against K-562 cell line. The recombinant DT386 and synthetic BR2 were used as the negative control in cytotoxicity assay. 
Results: The results of this study showed a significant reduction in survival of K-562 cells caused by DT386– BR2 as compared with BR2 and DT386 fragments. On the contrary, the flow-cytometry results showed apoptosis induction by DT386–BR2 after 12 h in a dose- and time-dependent manner. 
Conclusion: DT386–BR2 fusion protein can be used for further preclinical studies for determining its pharmacokinetic/ pharmacodynamic profiles and evaluating its anticancer efficacy in suitable animal models.