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Effects of FOXO3a Silencing or Over-expression on the Proliferation of
Huang Liya, Wang Fei, Cao Qing, Liu Fang, Sang Tiantian, Chen Shuyan*
Department of Geriatrics, Xinhua Hospital, Shanghai Jiao Tong University School of Medcine, Shanghai 200092, China
Abstract: To observe the effects of FOXO3a silencing or over-expression on the proliferation of endothelial
progenitor cells (EPCs)， the silent type Ad-shRNA-FOXO3a and the mutant type Ad-TM(triple mutant)-
FOXO3a recombinant adenovirus vectors were constructed. The CD133+/CD34+ EPCs from human cord blood
were isolated by density gradient centrifugation and fluorescence-activated cell sorting (FACS)， cultured in vitro
and then identified by immunofluorescent staining. The above-mentioned recombinant adenovirus vectors were
transfected into EPCs， the transfection efficiency and cell morphological changes were observed. The expression
changes of FOXO3a were detected by Western blot. Cells counting and MTT assay were used to assess the effects
of FOXO3a silencing or over-expression on the proliferation of EPCs. The recombinant adenovirus vectors AdshRNA-
FOXO3a and Ad-TM-FOXO3a were successfully constructed and transfected into EPCs from human cord
blood. The results of Western blot revealed that FOXO3a expression was obviously inhibited in transfected EPCs
with Ad-shRNA-FOXO3a， and was obviously increased in transfected EPCs with Ad-TM-FOXO3a. Cell morphological
changes， cells counting and MTT assay showed that the proliferation of EPCs was significantly promoted by effectively silencing of FOXO3a， and was significantly inhibited by over-expression of FOXO3a as compared with
control group. FOXO3a was involved in the regulation of cell proliferation of EPCs from human cord blood.
progenitor cells (EPCs)， the silent type Ad-shRNA-FOXO3a and the mutant type Ad-TM(triple mutant)-
FOXO3a recombinant adenovirus vectors were constructed. The CD133+/CD34+ EPCs from human cord blood
were isolated by density gradient centrifugation and fluorescence-activated cell sorting (FACS)， cultured in vitro
and then identified by immunofluorescent staining. The above-mentioned recombinant adenovirus vectors were
transfected into EPCs， the transfection efficiency and cell morphological changes were observed. The expression
changes of FOXO3a were detected by Western blot. Cells counting and MTT assay were used to assess the effects
of FOXO3a silencing or over-expression on the proliferation of EPCs. The recombinant adenovirus vectors AdshRNA-
FOXO3a and Ad-TM-FOXO3a were successfully constructed and transfected into EPCs from human cord
blood. The results of Western blot revealed that FOXO3a expression was obviously inhibited in transfected EPCs
with Ad-shRNA-FOXO3a， and was obviously increased in transfected EPCs with Ad-TM-FOXO3a. Cell morphological
changes， cells counting and MTT assay showed that the proliferation of EPCs was significantly promoted by effectively silencing of FOXO3a， and was significantly inhibited by over-expression of FOXO3a as compared with
control group. FOXO3a was involved in the regulation of cell proliferation of EPCs from human cord blood.
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