Rapamycin and 3-methyladenine regulate apoptosis and autophagy in bone-derived endothelial progenitor cells.
- Author:
Feng-rui LEI
1
;
Xiao-qiang LI
;
Hui LIU
;
Ren-da ZHU
;
Qing-you MENG
;
Jian-jie RONG
Author Information
- Publication Type:Journal Article
- MeSH: Adenine; analogs & derivatives; pharmacology; Animals; Apoptosis; drug effects; Autophagy; drug effects; Cell Cycle; drug effects; Cell Proliferation; drug effects; Cells, Cultured; Rats; Sirolimus; pharmacology
- From: Chinese Medical Journal 2012;125(22):4076-4082
- CountryChina
- Language:English
-
Abstract:
BACKGROUNDMammalian target of rapamycin (mTOR) is involved in a caspase independent form of programmed cell death called autophagy. The aim of this research was to investigate the effects of rapamycin and 3-methyladenine (3-MA) on autophagy, proliferation, apoptosis, and cell-cycle parameters of rat bone marrow-derived endothelial progenitor cells (EPCs).
METHODSMononuclear cells isolated from rat bone marrow were treated with rapamycin (0.01, 0.1, 1, or 10 µg/L) or 3-MA (1.25, 2.5, 5, or 10 mmol/L) for 24 hours. Expression of the autophagy marker protein LC3-II was analyzed by Western blotting. Apoptosis and cell-cycle progression were analyzed by flow cytometry. Cell proliferation was measured using the MTT assay.
RESULTSRapamycin treatment of EPCs induced apoptosis and autophagy and inhibited proliferation and cell-cycle progression in a dose-dependent manner. Treatment with 5 mmol/L 3-MA promoted cell proliferation; in contrast, treatment with 10 mmol/L 3-MA promoted apoptosis and induced S-phase arrest.
CONCLUSIONSRapamycin treatment of EPCs induced apoptosis and autophagy. Low concentrations of 3-MA had no significant effect on the proliferation and apoptosis of EPCs; The 5 mmol/L group promoted cell proliferation, but had no effect on the apoptosis; the 10 mmol/L group inhibited the proliferation and promoted apoptosis through the cell cycle.