[The effect of Syk recombinant adenovirus on phenotypic modulation in vascular smooth muscle cells].
OBJECTIVE
To construct Syk recombinant adenovirus with which vascular smooth muscle cells (VSMC) were infected, and to investigate the effect of Syk on the proliferation of VSMC.


METHODS
Syk adenovirus recombinant plasmid pDC315-GFP-Syk was constructed by using pDC315-GFP adenovirus vector system. The recombinant adenoviral vector (pDC315-GFP-Syk) was packaged and amplified in HEK 293 cells. The VSMC of rat pulmonary vascular smooth muscle cells were isolated and cultured in vitro and transfected with Syk adenovirus. The mRNA and protein express level of Syk, SM22alpha and alpha-SM-actin of rat pulmonary vascular smooth muscle cells were detected by RT-PCR and Western blot.


RESULTS
The recombinant adenovirus carrying rat Syk was constructed successfully. The titer of Syk adenovirus was 10(11) pfu/mL after purification. The expression of Syk mRNA (741638.70 +/- 35213.53) and protein (2.14 +/- 0.71) in VSMC transfected with Syk adenovirus was significantly higher (P < 0.01) than that of empty adenovirus after transfection at fifth day. The mRNA express of alpha-SM-actin (0.80 +/- 0.04) and SM22a (1.00 +/- 0.01) was significantly decreased (P < 0.05) in VSMC. Meanwhile, the protein express of alpha-SM-actin (0.61 +/- 0.10) and SM22alpha (0.18 +/- 0.06) was also significantly decreased (P < 0.01) in VSMC.


CONCLUSIONS
These data indicated that Syk played an important role in vascular remodeling by changing the phenotypes of VSMC.
Department of Cardiology, West China Second Hospital, Sichuan University, Chengdu 610041, China.
Zheng-xiang
ZX
Gao
Li-li
LL
Cao
Qin
Q
Luo
Li
L
Yu
Han-min
HM
Liu
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D005822:Genetic Vectors / Q000235:genetics*; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D010641:Phenotype; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis* / Q000235:genetics; D011651:Pulmonary Artery / Q000166:cytology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D000072377:Syk Kinase; D014162:Transfection
D047908:Intracellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D011505:Protein-Tyrosine Kinases; D000072377:Syk Kinase; C000606898:Syk protein, rat