Slfn1 inhibited the migration of endothelial progenitor cells in vitro.
- Author:
Lu ZHANG
1
;
Chunyan KUANG
2
;
Tianhe YANG
;
Qiang WU
;
Yang ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Cycle; Cell Cycle Proteins; physiology; Cell Division; Cell Movement; Cyclin D1; physiology; Down-Regulation; Endothelial Progenitor Cells; Flow Cytometry; In Vitro Techniques; RNA, Messenger; RNA, Small Interfering; Rats; Transfection; Up-Regulation
- From: Chinese Journal of Cardiology 2014;42(11):951-956
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the effect of Schlafen 1 (Slfn1) on the migration of endothelial progenitor cells (EPCs).
METHODSRat bone marrow derived EPCs were isolated and cultured. Ad-Slfn1, ShRNA-Slfn1, ShRNA-control and Ad-control were transfected into EPCs respectively. The mRNA expression of Slfn1 and Cyclin D1 was examined by reverse transcriptase-PCR, and their protein expression was detected by Western blot. The migration of EPCs was examined by a modified Boyden chamber assay.EPCs cell cycle was determined using flow cytometry analysis.
RESULTSForty-eight hours after ShRNA-Slfn1 transfection, the mRNA and protein expression of Slfn1 in EPCs was significantly down-regulated compared to ShRNA-control EPCs (P < 0.05). Transfection of Ad-Slfn1 reversed these changes.Overexpression of Slfn1 reduced the migration capacity of EPCs while the silencing of Slfn1 by shRNA-Slfn1 increased the migration capacity of EPCs.In addition, cell cycle was arrested at G1 phase in Slfn1 overexpression group while transfection of shRNA-Slfn1 reversed these responses.Interestingly, the mRNA and protein expression of Cyclin D1 was significantly up-regulated after shRNA-Slfn1 transfection compared to ShRNA-control group (all P < 0.05), but overexpression of Slfn1 reversed these results, suggesting Cyclin D1 was involved in regulating EPCs cell cycle via Slfn1 signaling.
CONCLUSIONSSlfn1 could reduce the migration capacity of EPCs via Cyclin D1 pathway.
