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Effects on proliferation and apoptosis of serum-induced rabbit VSMCs by adenovirus-mediated transfer of the Gax gene

ZHENG Hui,XUE Song,LIAN Feng,HU Zhen-lei,WANG Yong-yi Department of Cardiothoracic Surgery,Renji Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 200127,China  
AIM: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(P0.05,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(P0.05).CONCLUSION: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.
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