[Slfn1 inhibited the migration of endothelial progenitor cells in vitro]
Lu Zhang1, Chunyan Kuang2, Tianhe Yang
1Medical Examination Center, People's Hospital of Guizhou Province, Guiyang 550003, China.
Objective:
To explore the effect of Schlafen 1 (Slfn1) on the migration of endothelial progenitor cells (EPCs).
Methods:
Rat 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.
Results:
Forty-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.
Conclusions:
Slfn1 could reduce the migration capacity of EPCs via Cyclin D1 pathway.
