Treatment of human neuroblastoma cell line SH‐SY5Y with HSP27 siRNA tagged‐exosomes decreased differentiation rate into mature neurons

Heat shock proteins (HSPs) participate in the regulation of different cell activities in response to stimuli. By applying different strategies, the modulation of heat shock proteins is at the center of attention. Conventional delivery approaches are not fully encouraged due to cytotoxicity and immun...

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Bibliographic Details
Published in:Journal of cellular physiology 2019-11, Vol.234 (11), p.21005-21013
Main Authors: Shokrollahi, Elhameh, Nourazarian, Alireza, Rahbarghazi, Reza, Salimi, Leila, Karbasforush, Saeede, Khaksar, Majid, Salarinasab, Sadegh, Abhari, Alireza, Heidarzadeh, Morteza
Format: Article
Language:English
Subjects:
Assaying
Biomolecules
Bromodeoxyuridine
CD63 antigen
Cell differentiation
Cell proliferation
Cell viability
Cytotoxicity
differentiation
Differentiation (biology)
exosome
Exosomes
Flow cytometry
Heat shock proteins
HSP27
Hsp27 protein
human neuroblastoma cell line
Immunofluorescence
Immunogenicity
Neuroblastoma
Neuroblasts
Scanning electron microscopy
siRNA
Toxicity
Transfection
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Summary:Heat shock proteins (HSPs) participate in the regulation of different cell activities in response to stimuli. By applying different strategies, the modulation of heat shock proteins is at the center of attention. Conventional delivery approaches are not fully encouraged due to cytotoxicity and immunogenicity issues. Exosomes are touted as bio‐shuttles for delivery of distinct biomolecules inside the cells. Here, we aimed to HSP27 small interfering RNA (siRNA)‐tagged exosomes for the inhibition of Hsp27 in human neuroblastoma cell line SH‐SY5Y and explored differentiation into neuron‐like cells. Exosomes were isolated, characterized by scanning electron microscope (SEM) and CD63 then enriched with siRNA against Hsp27. Neuroblastoma cells were incubated with exosomes carrying siRNA for 48 hr. Exosome uptake was monitored by immunofluorescence assay. The cell viability and proliferation were analyzed using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide and bromodeoxyuridine/5‐bromo‐2′‐deoxyuridine incorporation assays. The ability of cells to form colonies was evaluated by clonogenic assay. The cell potential to express NeuN, a mature neuron factor, was studied by flow cytometry analysis. SEM showed the nano‐sized particles and a high level of CD63 after enrichment. Immunofluorescence imaging revealed an appropriate transfection rate in cell exposed to Hsp27 siRNA tagged exosomes. The cell viability and proliferation were reduced compared to cells received nude exosomes ( p 
ISSN:0021-9541
1097-4652
DOI:10.1002/jcp.28704