microRNA-1 gene delivery mediated by exosomes suppresses CAL-27 cell proliferation.
- Author:
Bao-Qin WU
1
;
Chun-Hui LI
1
;
Meng-Lian ZHANG
1
;
Min-Hai NIE
1
Author Information
- Publication Type:Journal Article
- Keywords: cell proliferation; exosomes; microRNA-1; oral squamous cell carcinoma
- MeSH: Cell Cycle; Cell Proliferation; Exosomes; HEK293 Cells; Humans; MicroRNAs
- From: West China Journal of Stomatology 2021;39(2):136-142
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVES:This study aims to construct endogenous exosomes abundantly loaded with miR-1 and investigate the role of exosome-mediated microRNA-1 (miR-1) delivery on CAL-27 cell proliferation.
METHODS:Exosomes secreted by miR-1-overexpressing HEK293 cells (miR1-EXO) were purified via ultracentrifugation and subjected to transmission electron microscopy, nanoparticle analysis, Western blot analysis, and quantitative polymerase chain reaction (qPCR). CAL-27 cells were cocultured with exosomes secreted by HEK293 cells (CON-EXO) and miR1-EXO and equivalent phosphate buffer saline. The intracellular transport of exosomes was measured by using immunofluorescence, the expression of miR-1 and its target gene MET were investigated via qPCR, CAL-27 cell proliferation was measured through MTT assay, and cell cycle state was determined by applying flow cytometry.
RESULTS:Electron microscopy revealed that miR1-EXO and CON-EXO were spherical or cup-shaped with an average diameter of approximately 110 nm. The well-known exosome markers CD9, Tsg101, and Alix were enriched. The expression of miR-1 in miR1-EXO was higher than that in CON-EXO (285.80±14.33 vs 1.00±0.06,
CONCLUSIONS:Exosomes secreted from miR1-EXO cells could load abundant miR-1. Exosomal miR-1 delivered into CAL-27 cells by using miR1-EXO suppressed the expression of MET mRNA and inhibited cell proliferation.
