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• W2023451161 endingPage "1587" @default.
• W2023451161 startingPage "1580" @default.
• W2023451161 abstract "Transfection efficiencies of non-viral gene delivery vectors commonly vary with cell type, owing to differences in proliferation rates and intracellular characteristics. Previous work demonstrated that the poly(diethylaminoethylmethacrylate) (PDEAEM)/Pluronic F127 pentablock copolymers exhibit transfection in vitro selectively in cancer cell lines as opposed to non-cancerous cell lines. This study continues the investigation of intracellular barriers to transfection using this vector in normal and cancer cell lines to understand the underlying mechanisms of the selectivity. Results from Part I of this investigation showed, using conjugated epidermal growth factor, that cellular uptake of these polyplexes is not a major barrier in these systems. Part II of this work continues the investigation into the other potential intracellular barriers, endosomal escape and nuclear entry, using a lysosomotropic agent chloroquine (CLQ), and a nuclear localization signal (NLS) SV40, respectively. Lack of effectiveness of NLS peptide in improving the transfection efficiency suggests that nuclear uptake might not be the major intracellular barrier using the pentablock copolymer vectors, or that the nuclear transport might not be primarily achieved through nuclear pores. However, inclusion of CLQ led to a dramatic enhancement in the level of gene expression, with an almost two orders of magnitude increase in expression seen in normal cell lines, compared with that the increase observed in cancer cell lines. The different lysosomal pH values in normal vs cancer cells was believed to cause the pentablock copolymer vectors to behave distinctly during transport through endocytic pathways, with greater loss of functional DNA occurring in normal cells containing more acidic endocytic vesicles in contrast to cancer cells with less acidic vesicles. Interestingly, CLQ introduced almost no enhancement in the transfection with the control vector ExGen which lacked selectivity of transfection. Exploiting intracellular differences between normal and cancer cells for gene delivery vector design offers a new paradigm to achieve transfection selectivity based on intracellular differences rather than conventional approaches involving vector modification using specific ligands for targeted delivery." @default.
• W2023451161 created "2016-06-24" @default.
• W2023451161 creator A5051350450 @default.
• W2023451161 creator A5072587945 @default.
• W2023451161 date "2011-04-01" @default.
• W2023451161 modified "2023-09-25" @default.
• W2023451161 title "The mechanism of selective transfection mediated by pentablock copolymers; Part II: Nuclear entry and endosomal escape" @default.
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• W2023451161 doi "https://doi.org/10.1016/j.actbio.2010.11.033" @default.
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• W2023451161 hasPublicationYear "2011" @default.
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