D. Goukassian et al., Overexpression of p27(Kip1) by doxycycline-regulated adenoviral vectors inhibits endothelial cell proliferation and migration and impairs angiogenesis, FASEB J, 15(11), 2001, pp. 1877-1885
Formation of new blood vessels in the adult animal (i.e., angiogenesis) is
an important event for tissue repair and for tumor growth and metastasis. A
ngiogenesis involves the migration and proliferation of endothelial cells.
We have investigated the role of the growth suppressor p27(Kip1) (p27) on e
ndothelial cell function in vitro and angiogenesis in vivo. We have generat
ed Ad-TetON, a replication-deficient adenovirus that constitutively express
es the reverse tet-responsive transcriptional activator, and Ad-TRE-p27, wh
ich drives expression of p27 under the control of the tet response element.
Western blot analysis demonstrated doxycycline-dependent overexpression of
p27 in human umbilical vein endothelial cells (HUVECs) coinfected with Ad-
TetON and Ad-TRE-p27, which resulted in a marked inhibition of DNA replicat
ion and cell migration in vitro. Inducible overexpression of p27 in culture
d HUVECs inhibited the formation of tubelike structures and, when applied i
n a murine model of hind limb ischemia, reduced hind limb blood flow recove
ry and capillary density. These findings thus underscore a novel role of p2
7 in regulating endothelial cell migration in vitro and angiogenesis in viv
o, suggesting a novel anti-angiogenic therapy based on inducible p27 overex
pression.