Abstract
Objective
In this study, transfection efficiency of human papillomavirus (HPV) E7 DNA and protein constructs into HEK-293T normal cell line, and A549 and TC-1 tumor cell lines was evaluated by four delivery systems including supercharge GFP, hPP10 cell penetrating peptide, TurboFect and Lipofectamine using fluorescence microscopy and flow cytometry.
Results
The results indicated that Lipofectamine 2000 and TurboFect produced more effective transfection for GFP and E7-GFP DNA constructs in HEK-293T cells compared to in A549 and TC-1 cells (p < 0.05). In contrast, the supercharge GFP was efficient for E7 DNA and E7 protein delivery in both normal cell (~ 83.94 and ~ 77.01% for HEK-293T), and cancer cells (~ 71.69 and ~ 67.19% for TC-1, and ~ 73.86 and ~ 67.49% for A549), respectively. Indeed, in these cell lines, transfection efficiency by +36 GFP reached ~ 60–80%. Moreover, the hPP10 produced the best transfection result for E7-GFP protein in HEK-293T cells (~ 63.66%) compared to TurboFect (~ 32.95%); however, the efficiency level of hPP10 was only ~ 17.51 and ~ 16.36% in TC-1 and A549 cells.
Conclusions
Our data suggested that the supercharge GFP is the most suitable transfection vehicle for DNA and protein delivery into TC-1 and A549 tumor cell lines compared to other carriers.
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Acknowledgements
Financial support of this work was provided by Virology Research Center, Shahid Beheshti University of Medical Sciences, and Pasteur Institute of Iran.
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Supplementary data
Physiochemical characterization and stability analysis of the +36GFP/DNA nanoparticles: A) Representative gel retardation assay of +36 GFP complexed with pcDNA-E7 at different N/P ratios (GFP: E7DNA); Lane 1: naked plasmid DNA as a control (pcDNA-E7), Lane 2: N/P = 1:1, Lane 3: N/P = 2:1, Lane 4: N/P = 5:1, Lane 5: N/P = 10:1, and Lane 6: N/P = 20:1. The DNA complexed with GFP that was not able to migrate into the gels was observed at an N/P ratio of 5:1; B) Stability analysis of GFP-based nanoparticles against DNase I; Lane 1: naked plasmid DNA with DNase, Lane 2: naked plasmid DNA without DNase, and Lane 3: N/P = 10:1.
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Shahbazi, S., Haghighipour, N., Soleymani, S. et al. Delivery of molecular cargoes in normal and cancer cell lines using non-viral delivery systems. Biotechnol Lett 40, 923–931 (2018). https://doi.org/10.1007/s10529-018-2551-2
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DOI: https://doi.org/10.1007/s10529-018-2551-2