[Secretory expression of PR39 following adeno-associated viral-encoding fusion gene transfer induces angiogenesis in hypoxia chick embryo].
OBJECTIVE
To investigate the impact of AAV-encoding NT4-TAT-His-PR39 fusion gene expression on HIF-1alpha level in ECV304 cultured under hypoxic condition (1%O(2)) and on angiogenesis in hypoxic chick embryo.


METHODS
PR39 cDNA was connected with NT4, TAT, 6 x His cDNA by molecular biology methods. The recombinant AAV vector was obtained by three plasmid co-transfection in 293 cells. Then ECV304 were respectively infected with AAV-NT4-TAT-His-PR39, 6 x His expression and HIF-1alpha level in ECV304 were detected by immunocytochemistry. The chicken embryos were randomized into the AAV-PR39, EV and PBS groups (n = 10 each) subject to hypoxia (5%O(2), n = 15) or normoxia environments (n = 15), the vessel density of the chicken chorioallantoic membrane (CAM) were measured by Image Pro Plus (IPP) software.


RESULTS
The expression of 6 x His protein was detected in AAV-PR39 infected ECV304 cells. HIF-1alpha protein activity was significantly increased in AAV-PR39 infected ECV304 underwent hypoxia compared to PBS and non-infected ECV304 groups (P < 0.05). The vessel density of chicken CAM in hypoxia environment but not in normoxia environment was also significantly higher in AAV-PR39 group than in EV group and PBS group (all P < 0.05).


CONCLUSIONS
AAV-encoding NT4-TAT-His-PR39 fusion gene expression significantly increased HIF-1alpha level in ECV304 exposed to hypoxia and promoted angiogenesis in hypoxic chicken embryo.
Hao
Yue-wen
YW
Department of Radiology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Sun
Li-jun
LJ
Liu
Ying
Y
Wang
Quan-ying
QY
Yang
Guang-xiao
GX
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics*; D002460:Cell Line; D002642:Chick Embryo; D000229:Dependovirus / Q000235:genetics; D050939:Gene Fusion; D018014:Gene Transfer Techniques; D005814:Genes, Viral; D000860:Hypoxia / Q000235:genetics*; D018919:Neovascularization, Physiologic / Q000235:genetics*
D023181:Antimicrobial Cationic Peptides; C072212:PR 39