| 조현주¹, 김광일² ,한주용¹ ,전수인¹ ,채인호² ,김효수¹,손대원¹ ,오병희¹ ,이명묵¹ ,박영배¹ ,최윤식¹ |
Backgrounds: β-catenin has been known to be a transcriptional regulator of several genes that are involved in proliferation and survival. However, little information is available regarding a potential role for β-catenin in angiogenesis. The goal of this study was to examine the role of β-catenin on endothelial cell(EC)s, and on the process of angiogenesis.
Methods & Results: Adenoviral mediated β-catenin gene transfer resulted in enhanced serum stimulated EC proliferation (WT: GFP= 204.9±15.6: 100.0±28.3%, p<0.05, by WST-1 assay) and protected ECs from serum-deprivation-induced apoptosis (sub-2N population of WT: GFP= 16.0: 42.3%, p<0.05, by FACS analysis). Furthermore, the capillary forming capabilities of ECs were augmented as shown by tube formation assay. After transduction of β-catenin into HUVECs, cyclin E2, which operates during the G1/S phase progression, was increased. In contrast, VEGF and cyclin D1 expression were increased in skeletal muscle. Increased expression of VEGF in skeletal muscle promoted ECs proliferation which was blocked by anti-VEGF neutralizing antibody. The bioactivity of β-catenin gene transfer in vivo was examined in a murine model of hindlimb ischemia. To impair angiogenesis in response to hindlimb ischemia, mouse was fed with 2% high-cholesterol diet. At 2 weeks of the dietary intervention, unilateral limb ischemia was surgically induced in all animals. Adenovirus containing either the β-catenin (n=10) or GFP gene (n=10) were injected into the adductor muscle of mouse immediately after femoral artery removal (1 ´ 109 pfu per mouse, total 40ul divided into 4 sites). Intramuscular injection of β-catenin resulted in significant increase of blood perfusion and capillary density associated with enhanced VEGF expression. Conclusions: These data suggest that the effect of β-catenin on ECs may be modulation of the cell cycle and proliferation by inducing VEGF expression. Furthermore, adenovirus-mediated β-catenin gene transfer may be a promising genetic modification to improve EC function and proliferation, and therapeutic angiogenesis.
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