Adipocyte-derived leucine aminopeptidase suppresses angiogenesis in human endometrial carcinoma via renin-angiotensin system

Clin Cancer Res. 2003 Dec 15;9(17):6497-503.

Abstract

Purpose: Angiotensin (Ang) II was reported to induce vascular endothelial growth factor (VEGF) expression in various cells. Adipocyte-derived leucine aminopeptidase (A-LAP) is a novel member of the M1 family of zinc metallopeptidases. Enzymatic characterization demonstrated that A-LAP hydrolyzes Ang II. This study examined the role of A-LAP in angiogenesis of human endometrial carcinoma.

Experimental design: We investigated whether Ang II induces VEGF expression in human endometrial carcinoma cells. To investigate the possible function of A-LAP in angiogenesis of endometrial carcinoma, we transfected A-LAP cDNA into HEC-1A cells, showing the lowest expression of A-LAP.

Results: In the present study, we showed that Ang II enhanced VEGF expression in a dose-dependent manner in endometrial carcinoma cells (HEC-1A cells). Overexpression of A-LAP attenuated Ang II-induced VEGF expression in HEC-1A cells. In addition, Human umbilical vascular endothelial cell migration was increased in conditioned media from Ang II-treated wild-type cells, but not in conditioned media from Ang II-treated A-LAP-overexpressing cells (HEC-1A-A-LAP cells). In an in vivo study, we showed that A-LAP overexpression in endometrial carcinoma cells results in a reduction of VEGF immunoreactivity and the number of blood vessels within tumors.

Conclusions: Our study demonstrates that it is feasible to overexpress Ang II-degrading enzymes in cultured cells and that this overexpression attenuated some effects of exogenous and endogenous Ang II. These experiments are a first step toward the development of novel strategies to selectively antagonize locally generated Ang II and suppress VEGF-induced angiogenesis in endometrial carcinoma.

MeSH terms

  • Adipocytes / metabolism*
  • Angiotensin II / metabolism
  • Animals
  • Blotting, Western
  • Carcinoma / metabolism*
  • Cell Line, Tumor
  • Cell Movement
  • Cells, Cultured
  • Culture Media, Conditioned / pharmacology
  • DNA, Complementary / metabolism
  • Dose-Response Relationship, Drug
  • Endometrial Neoplasms / metabolism*
  • Endothelium, Vascular / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Immunohistochemistry
  • Leucyl Aminopeptidase / metabolism*
  • Mice
  • Mice, Nude
  • Neovascularization, Pathologic*
  • Plasmids / metabolism
  • Renin-Angiotensin System
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transfection
  • Zinc / metabolism

Substances

  • Culture Media, Conditioned
  • DNA, Complementary
  • Angiotensin II
  • Leucyl Aminopeptidase
  • Zinc