Intracellular Delivery of Antisense Peptide Nucleic
Acid by Fluorescent Mesoporous Silica Nanoparticles
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Abstract
In order to overcome poor cell permeability
of antisense peptide
nucleic acid (PNA), a fluorescent mesoporous silica nanoparticle (MSNP)
carrier was developed to successfully deliver antisense PNA into cancer
cells for effective silence of B-cell lymphoma 2 (Bcl-2) protein expression <i>in vitro</i>. First, fluorescent MSNP functionalized with disulfide
bond bridged groups was fabricated and characterized. Antisense and
negative control PNAs were synthesized and further conjugated with
fluorescent dye cyanine 5. Then, the PNAs were covalently connected
with fluorescent MSNP via amidation between amino group of PNAs and
carboxylic acid group on the MSNP surface. High intracellular concentration
of glutathione serves as a natural reducing agent, which could cleave
the disulfide bond to trigger the PNA release <i>in vitro</i>. Confocal laser scanning microscopy studies prove that PNA conjugated
MSNP was endocytosed by HeLa cancer cells, and redox-controlled intracellular
release of antisense PNA from fluorescent MSNP was successfully achieved.
Finally, effective silencing of the Bcl-2 protein expression induced
by the delivered antisense PNA into HeLa cells was confirmed by Western
blot assay