Suicide gene therapy-mediated purine nucleoside phosphorylase/fludarabine system for in vitro breast cancer model with emphasis on evaluation of vascular endothelial growth factor promoter efficacy
- 1 March 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in 3 Biotech
- Vol. 11 (3), 1-14
- https://doi.org/10.1007/s13205-021-02692-0
Abstract
In this study, a suicide gene therapy approach was optimized by a non-viral polyplex system based on pEGFP-N1 vector harboring purine nucleoside phosphorylase gene conducted by vascular endothelial growth factor promoter for an in vitro breast cancer model (4T1 cell line). The VEGF promoter and purine nucleoside phosphorylase gene were cloned into the vector from the source of 4T1 and E. coli genomic DNA, respectively. A gene construct was developed by replacing VEGF promoter instead of CMV promoter in pEGFP-N1vector. PNP gene was integrated in to the multiple cloning site of the obtained vector. On the other hand, a construct from pEGFP-N1 harboring PNP gene under the control of the original CMV promoter was developed. The transfection method using cationic polymer was optimized based on N/P ratio, cell cytotoxicity, polyplex size, zeta potential and the green fluorescent protein (GFP) expression by fluorescent microscopy and flowcytometry. Also, the effect of hypoxia condition induced by 0.5 mM H2O2 on the promoter efficiency was investigated. The results showed that the performed gene delivery system is capable of the gene transfection to more than 30% of the cancer cells with both VEGF-PNP-pEGFP-N1 and PNP-pEGFP-N1 plasmids. The hypoxia condition did not show a significant effect on the VEGF promoter. But, it revealed that bystander effect can improve the efficacy of this system and reduce drug IC50 to 2 and fourfold for plasmids VEGF-PNP-pEGFP-N1 and PNP-pEGFP-N1, respectively. These results showed that the bystander effect could almost compensate the low efficiency of non-viral gene delivery systems. We suggest that the tumor-specific gene expression system mediated by the VEGF promoter can be especially useful in the present model of breast cancer gene therapy.Funding Information
- Zanjan University of Medical Sciences (A-12-848-16)
This publication has 46 references indexed in Scilit:
- Purine Nucleoside Phosphorylase Targeted by Annexin V to Breast Cancer Vasculature for Enzyme Prodrug TherapyPLOS ONE, 2013
- Porous Chitosan Scaffolds with Embedded Hyaluronic Acid/Chitosan/Plasmid-DNA Nanoparticles Encoding TGF-β1 Induce DNA Controlled Release, Transfected Chondrocytes, and Promoted Cell ProliferationPLOS ONE, 2013
- Suicide gene therapy for hepatocellular carcinoma cells by survivin promoter-driven expression of the herpes simplex virus thymidine kinase geneOncology Reports, 2013
- Drying of a plasmid containing formulation: chitosan as a protecting agentDARU Journal of Pharmaceutical Sciences, 2012
- Cationic lipid-coated PEI/DNA polyplexes with improved efficiency and reduced cytotoxicity for gene delivery into mesenchymal stem cellsInternational Journal of Nanomedicine, 2012
- Novel pH-sensitive cationic lipids with linear ortho ester linkers for gene deliveryEuropean Journal of Medicinal Chemistry, 2012
- Cellular Cytoskeleton Dynamics Modulates Non-Viral Gene Delivery through RhoGTPasesPLOS ONE, 2012
- FOXO3a represses VEGF expression through FOXM1-dependent and -independent mechanisms in breast cancerOncogene, 2011
- Targeted Therapies in Epithelial Ovarian CancerCancers, 2010
- Tumor-specific adenovirus-mediated PUMA gene transfer using the survivin promoter enhances radiosensitivity of breast cancer cells in vitro and in vivoBreast Cancer Research and Treatment, 2008