[tMfn2 gene transfer promotes vascular smooth-muscle cells apoptosis via activation of the mitochondrial cell death pathway].
OBJECTIVE
To investigate the effect of tMfn2 gene transfer on promoting the apoptosis of vascular smooth-muscle cells (VSMCs).


METHODS
VSMCs were infected by adenovirus-mediated tMfn2 (Adv-tMfn2) or adenovirus-mediated Mfn2 (Adv-Mfn2). FACS analysis, cell death ELISA and TUNEL staining were used to investigate the role of tMfn2 and Adv-Mfn2 gene transfer on VSMCs apoptosis. Western blot was used to analyze the protein expression of p-Akt, Bcl-2, Bax and cleaved caspase-9.


RESULTS
FACS and ELISA results showed that tMfn2 was superior to Mfn2 in promoting VSMCs apoptosis and tMfn2 gene transfer induced VSMCs apoptosis in a time-dependent manner (P < 0.01). TUNEL staining evidenced that there were more positive-apoptotic VSMCs in tMfn2 group than that in Mfn2 group (P < 0.01). The protein expressions of phosphorylated Akt and Bcl-2 were significantly decreased, whereas Bax and cleaved caspase-9 protein expressions were significantly upregulated in tMfn2-transfected VSMCs.


CONCLUSIONS
tMfn2 transfer promoted apoptosis of VSMCs in a time dependent manner via the PI3K-Akt signaling pathway and mitochondrial apoptotic pathway.
Zhao
Li
L
Department of Cardiology, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhou
Wei
W
Wang
Si-kun
SK
Liao
Hua
H
Zhang
Wen-juan
WJ
Chen
Guang-hui
GH
Guo
Xiao-mei
XM
D000256:Adenoviridae; D000818:Animals; D017209:Apoptosis; D053453:Caspase 9 / Q000378:metabolism; D002478:Cells, Cultured; D020558:GTP Phosphohydrolases; D008565:Membrane Proteins / Q000235:genetics*; D008928:Mitochondria / Q000378:metabolism*; D051336:Mitochondrial Membranes / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000235:genetics*; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D014162:Transfection; D051028:bcl-2-Associated X Protein / Q000378:metabolism
C494828:Bax protein, rat; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D051057:Proto-Oncogene Proteins c-akt; C506435:Casp9 protein, rat; D053453:Caspase 9; D020558:GTP Phosphohydrolases; C489710:Mfn2 protein, rat