[Effects on proliferation and apoptosis of serum-induced rabbit VSMCs by adenovirus-mediated transfer of the Gax gene].
OBJECTIVE
To investigate the effect of a recombinant replication-incompetent Ad5-hGax vector on the proliferation and apoptosis of serum-induced rabbit vascular smooth muscle cells (VSMCs) in vitro,  and to provide an important support that Gax would be an optimal gene for gene therapy in vein graft failure.


METHODS
The Ad5-hGax vector was infected into rabbit VSMCs,  then the protein level of rabbit VSMCs was detected at 1 d,  3 d,  5 d by Western blot,  respectively. MTT was applied to observe the inhibitory effects of overexpressed hGax on serum-induced rabbit VSMCs. the apoptosis of serum-induced rabbit VSMCs was measured by using flow cytometry in 72 h after transfection.


RESULTS
The protein expression of human Gax in the Ad5-hGax transfected cells on 1st day,  3rd day,  5th day was determined; MTT showed that the proliferation of serum-induced rabbit VSMCs was significantly inhibited in Ad5-hGax group at 48 h,  72 h and 96 h (P<0.05,  respectively);  After transfecting for 72 h,  flow cytometry analysis showed that the number of the apoptosis cells was increased in serum-induced Rabbit VSMCs of Ad5-hGax group (P<0.05).


CONCLUSIONS
Overexpressed hGax could inhibit the proliferation of serum-induced Rabbit VSMCs and induce their apoptosis. These findings carry significant implications for adenovirus vector-based Gax gene therapies for vein graft failure.
Department of Cardiothoracic Surgery, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200127, China. 123123zhenghui@163.com
Hui
H
Zheng
Song
S
Xue
Feng
F
Lian
Zhen-lei
ZL
Hu
Yong-yi
YY
Wang
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D049109:Cell Proliferation; D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D011817:Rabbits
D018398:Homeodomain Proteins; C484193:MEOX2 protein, human