Low Electric Treatment activates Rho GTPase via Heat Shock Protein 90 and Protein Kinase C for Intracellular Delivery of siRNA.

Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE

Original Publication: London : Nature Publishing Group, copyright 2011-

Electricity*; HSP90 Heat-Shock Proteins/*metabolism ; Intracellular Space/*metabolism ; Protein Kinase C/*metabolism ; RNA, Small Interfering/*administration & dosage ; rho GTP-Binding Proteins/*metabolism; Actin Cytoskeleton/metabolism ; Animals ; Calcium/metabolism ; Isotope Labeling ; Macromolecular Substances/metabolism ; Mice ; Models, Biological ; NIH 3T3 Cells ; Phosphorylation ; Up-Regulation

Low electric treatment (LET) promotes intracellular delivery of naked siRNA by altering cellular physiology. However, which signaling molecules and cellular events contribute to LET-mediated siRNA uptake are unclear. Here, we used isobaric tags in relative and absolute quantification (iTRAQ) proteomic analysis to identify changes in the levels of phosphorylated proteins that occur during cellular uptake of siRNA promoted by LET. iTRAQ analysis revealed that heat shock protein 90 (Hsp90)α and myristoylated alanine-rich C-kinase substrate (Marcks) were highly phosphorylated following LET of NIH 3T3 cells, but not untreated cells. Furthermore, the levels of phosphorylated Hsp90α and protein kinase C (PKC)γ were increased by LET both with siRNA and liposomes having various physicochemical properties used as model macromolecules, suggesting that PKCγ activated partly by Ca ²⁺ influx as well as Hsp90 chaperone function were involved in LET-mediated cellular siRNA uptake. Furthermore, LET with siRNA induced activation of Rho GTPase via Hsp90 and PKC, which could contribute to cellular siRNA uptake accompanied by actin cytoskeleton remodeling. Collectively, our results suggested that LET-induced Rho GTPase activation via Hsp90 and PKC would participate in actin-dependent cellular uptake of siRNA.

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0 (HSP90 Heat-Shock Proteins)
0 (Macromolecular Substances)
0 (RNA, Small Interfering)
EC 2.7.11.13 (Protein Kinase C)
EC 3.6.5.2 (rho GTP-Binding Proteins)
SY7Q814VUP (Calcium)

Date Created: 20190313 Date Completed: 20201020 Latest Revision: 20210109

20250114

PMC6412017

10.1038/s41598-019-40904-z

30858501