Objective        To construct an adenovirus vector of transcription factor E2-2 gene and investigate whether the adenovirus vector carrying E2-2 gene affects the expression of inhibitor of DNA binding/differentiation (ID1) in endothelial progenitor cells (EPCs).       Methods        Mur

ine EPCs of bone marrow were isolated

cultured and identified. cDNA fragment encoding murine E2-2 gene was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR). The PCR amplified fragment was first cloned into pTG19-T vector

and then sub-cloned into the shuttle vector pAdTrack-CMV in order to construct the recombinant plasmid pAdTrack/E2-2

which homologously recombined with the adenoviral backbone vectors Adeasy-1 to generate recombinant adenoviral plasmid Ad/E2-2. The recombinant adenoviruses Ad/E2-2 were generated by transfecting the recombinant adenoviral DNA into 293 cells

and then employed to infect EPCs. The expression of enhanced green fluorescent protein (EGFP) in the transfected EPCs were observed by invert microscopy. The growth and proliferation of transfected EPCs were tested

via

Cell Count Kit-8 (CCK-8)

and the expression of E2-2 and ID1 in transfected EPCs were examined by RT-PCR and Western blotting respectively

and the expression of ID1 was semi-quantitated.       Results        Murine EPCs of bone marrow were isolated

cultured and identified. Murine E2-2 gene was successfully cloned. Adenoviral virus particle which possesses infectious competent was produced from Ad/E2-2. The results of CCK-8 demonstrated that the growth and proliferation of EPCs

which over-expressed E2-2

were slower than the control cells

especially from 48 h after transfection (

P

0.01). The results of RT-PCR

Western blotting and quantitative analysis revealed that E2-2 down-regulated the expression of ID1 in EPCs when compared with control cells (

P

0.01).       Conclusion        The recombinant adenoviral vector of E2-2 is constructed successfully. The vector containing E2-2 gene prevents the EPCs from growth and proliferation and down-regulates the expression of ID1 in the cells.