Delivery of molecular cargoes in normal and cancer cell lines using non-viral delivery systems
- 69 Downloads
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.
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.
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.
KeywordsHuman papillomavirus E7 protein Delivery system hPP10 +36 GFP TuboFect Lipofectamine
Financial support of this work was provided by Virology Research Center, Shahid Beheshti University of Medical Sciences, and Pasteur Institute of Iran.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
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.
- Kadkhodayan S, Sadat SM, Irani S, Fotouhi F, Bolhassani A (2016) Generation of GFP native protein for detection of its intracellular uptake by cell-penetrating peptides. Folia Biol 62:103–109Google Scholar
- Margie L (2015) The discovery and characterization of endosomal escape enhancing compounds to improve protein delivery efficacy. Doctoral dissertation, Harvard University, Graduate School of Arts & Sciences pp 1–115Google Scholar
- Shahbazi S, Bolhassani A, Arashkia A, Sadroddiny E (2018) Generation of the fluorescent HPV16 E7 protein for detection of delivery in vitro. Protein Pept Lett. https://doi.org/10.2174/0929866525666180115123620 (in press)
- Thompson DB, Cronican JJ, Liu DR (2012) Engineering and identifying supercharged proteins for macromolecule delivery into mammalian cells. Methods Enzymol 503:293–319. https://doi.org/10.1016/B978-0-12-396962-0.00012-4 CrossRefPubMedPubMedCentralGoogle Scholar
- Tong H, Liu H, Wang Y, Yang F, Shi Q, Fernandes JC et al (2014) A novel in vitro system for intracellular delivery of non-viral DNA. J Orthop Transl 2:157–164Google Scholar
- Wang H, Ma JL, Yang YG, Song Y, Wu J, Qin YY et al (2016) Efficient therapeutic delivery by a novel cell-permeant peptide derived from KDM4A protein for anti-tumor and anti-fibrosis. Oncotarget 7(31):1–16Google Scholar