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Induction of the angiogenic phenotype by Hox D3.

Boudreau N, Andrews C, Srebrow A, Ravanpay A, Cheresh DA - J. Cell Biol. (1997)

Bottom Line: Expression of Hox D3, in the absence of bFGF, resulted in enhanced expression of integrin alphavbeta3 and uPA.In fact, sustained expression of Hox D3 in vivo on the chick chorioallantoic membrane retained EC in this invasive state and prevented vessel maturation leading to vascular malformations and endotheliomas.Therefore, Hox D3 regulates EC gene expression associated with the invasive stage of angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Vascular Biology, The Scripps Research Institute, La Jolla, California 92037, USA. nboudrea@nsc.vcu.edu

ABSTRACT
Angiogenesis is characterized by distinct phenotypic changes in vascular endothelial cells (EC). Evidence is provided that the Hox D3 homeobox gene mediates conversion of endothelium from the resting to the angiogenic/invasive state. Stimulation of EC with basic fibroblast growth factor (bFGF) resulted in increased expression of Hox D3, integrin alphavbeta3, and the urokinase plasminogen activator (uPA). Hox D3 antisense blocked the ability of bFGF to induce uPA and integrin alphavbeta3 expression, yet had no effect on EC cell proliferation or bFGF-mediated cyclin D1 expression. Expression of Hox D3, in the absence of bFGF, resulted in enhanced expression of integrin alphavbeta3 and uPA. In fact, sustained expression of Hox D3 in vivo on the chick chorioallantoic membrane retained EC in this invasive state and prevented vessel maturation leading to vascular malformations and endotheliomas. Therefore, Hox D3 regulates EC gene expression associated with the invasive stage of angiogenesis.

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Effects of Hox D3 sense and antisense expression on  EC adhesion and proliferation. (A) Adhesion to microtitre wells  coated with 10 μg/ml fibrinogen by EC transfected with Hox D3  sense (▪) or antisense (▨ ▨ ▨ ) expression vectors. Adhesion after 30  min was assessed by absorbance at 600 nm and expressed as a  percentage of adhesion displayed by control-transfected EC (n =  3). *P < 0.05. (B) BrdU incorporation in control-transfected EC  (□), Hox D3-overexpressing cells (▪), or cells expressing antisense against Hox D3 (▨ ▨ ▨ ). Cells were labeled for 4 or 12 h with  10 mM BrdU. The percentage of cells staining positive for BrdU  was assessed by counting at least six different fields containing a  total of at least 400 cells each.
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Figure 3: Effects of Hox D3 sense and antisense expression on EC adhesion and proliferation. (A) Adhesion to microtitre wells coated with 10 μg/ml fibrinogen by EC transfected with Hox D3 sense (▪) or antisense (▨ ▨ ▨ ) expression vectors. Adhesion after 30 min was assessed by absorbance at 600 nm and expressed as a percentage of adhesion displayed by control-transfected EC (n = 3). *P < 0.05. (B) BrdU incorporation in control-transfected EC (□), Hox D3-overexpressing cells (▪), or cells expressing antisense against Hox D3 (▨ ▨ ▨ ). Cells were labeled for 4 or 12 h with 10 mM BrdU. The percentage of cells staining positive for BrdU was assessed by counting at least six different fields containing a total of at least 400 cells each.

Mentions: The Hox D3-mediated changes in expression of integrin β3 mRNA also resulted in corresponding changes in surface expression of functional αvβ3 integrin. We compared the ability of EC overexpressing Hox D3 or antisense against Hox D3 to adhere to fibrinogen, a ligand for αvβ3 on endothelial cells (Cheresh et al., 1989). EC expressing antisense against Hox D3 displayed significantly reduced capacity to adhere to fibrinogen, compared to control cells (Fig. 3 A), whereas overexpressing Hox D3 increased EC adhesion to fibrinogen (Fig. 3 A). Treatment of cells with LM609, a function-blocking antibody against αvβ3 (Cheresh, 1987), resulted in a complete inhibition of the fibrinogen-mediated attachment of these cells (data not shown). Thus, Hox D3 regulates the functional expression of αvβ3 on endothelial cells.


Induction of the angiogenic phenotype by Hox D3.

Boudreau N, Andrews C, Srebrow A, Ravanpay A, Cheresh DA - J. Cell Biol. (1997)

Effects of Hox D3 sense and antisense expression on  EC adhesion and proliferation. (A) Adhesion to microtitre wells  coated with 10 μg/ml fibrinogen by EC transfected with Hox D3  sense (▪) or antisense (▨ ▨ ▨ ) expression vectors. Adhesion after 30  min was assessed by absorbance at 600 nm and expressed as a  percentage of adhesion displayed by control-transfected EC (n =  3). *P < 0.05. (B) BrdU incorporation in control-transfected EC  (□), Hox D3-overexpressing cells (▪), or cells expressing antisense against Hox D3 (▨ ▨ ▨ ). Cells were labeled for 4 or 12 h with  10 mM BrdU. The percentage of cells staining positive for BrdU  was assessed by counting at least six different fields containing a  total of at least 400 cells each.
© Copyright Policy
Related In: Results  -  Collection

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Figure 3: Effects of Hox D3 sense and antisense expression on EC adhesion and proliferation. (A) Adhesion to microtitre wells coated with 10 μg/ml fibrinogen by EC transfected with Hox D3 sense (▪) or antisense (▨ ▨ ▨ ) expression vectors. Adhesion after 30 min was assessed by absorbance at 600 nm and expressed as a percentage of adhesion displayed by control-transfected EC (n = 3). *P < 0.05. (B) BrdU incorporation in control-transfected EC (□), Hox D3-overexpressing cells (▪), or cells expressing antisense against Hox D3 (▨ ▨ ▨ ). Cells were labeled for 4 or 12 h with 10 mM BrdU. The percentage of cells staining positive for BrdU was assessed by counting at least six different fields containing a total of at least 400 cells each.
Mentions: The Hox D3-mediated changes in expression of integrin β3 mRNA also resulted in corresponding changes in surface expression of functional αvβ3 integrin. We compared the ability of EC overexpressing Hox D3 or antisense against Hox D3 to adhere to fibrinogen, a ligand for αvβ3 on endothelial cells (Cheresh et al., 1989). EC expressing antisense against Hox D3 displayed significantly reduced capacity to adhere to fibrinogen, compared to control cells (Fig. 3 A), whereas overexpressing Hox D3 increased EC adhesion to fibrinogen (Fig. 3 A). Treatment of cells with LM609, a function-blocking antibody against αvβ3 (Cheresh, 1987), resulted in a complete inhibition of the fibrinogen-mediated attachment of these cells (data not shown). Thus, Hox D3 regulates the functional expression of αvβ3 on endothelial cells.

Bottom Line: Expression of Hox D3, in the absence of bFGF, resulted in enhanced expression of integrin alphavbeta3 and uPA.In fact, sustained expression of Hox D3 in vivo on the chick chorioallantoic membrane retained EC in this invasive state and prevented vessel maturation leading to vascular malformations and endotheliomas.Therefore, Hox D3 regulates EC gene expression associated with the invasive stage of angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Vascular Biology, The Scripps Research Institute, La Jolla, California 92037, USA. nboudrea@nsc.vcu.edu

ABSTRACT
Angiogenesis is characterized by distinct phenotypic changes in vascular endothelial cells (EC). Evidence is provided that the Hox D3 homeobox gene mediates conversion of endothelium from the resting to the angiogenic/invasive state. Stimulation of EC with basic fibroblast growth factor (bFGF) resulted in increased expression of Hox D3, integrin alphavbeta3, and the urokinase plasminogen activator (uPA). Hox D3 antisense blocked the ability of bFGF to induce uPA and integrin alphavbeta3 expression, yet had no effect on EC cell proliferation or bFGF-mediated cyclin D1 expression. Expression of Hox D3, in the absence of bFGF, resulted in enhanced expression of integrin alphavbeta3 and uPA. In fact, sustained expression of Hox D3 in vivo on the chick chorioallantoic membrane retained EC in this invasive state and prevented vessel maturation leading to vascular malformations and endotheliomas. Therefore, Hox D3 regulates EC gene expression associated with the invasive stage of angiogenesis.

Show MeSH
Related in: MedlinePlus