title: Extra cellular matrix derived metabolite regulates angiogenesis by FasL mediated apoptosis.
abstract: OBJECTIVE
  Antiangiogenic treatments are beginning to give promising outcomes in many vascular diseases including tumor angiogenesis. In this current study the antiangiogenic and pro-apoptotic actions of α1(IV)NC1 and its N- and C- peptides α1S1(IV)NC1, α1S2(IV)NC1 were investigated in-vitro and in-vivo.
  
  
  METHODS
  Endothelial cells (ECs) were treated with α1(IV)NC1, α1S1(IV)NC1, α1S2(IV)NC1 and in-vitro proliferation, migration, tube formation and apoptotic assays were executed. FasL, Fas, Caspase-8, -3 and PARP activations were studied using immunoblotting analysis using specific antibodies. Also the in-vivo antiangiogenic and pro-apoptotic effects were tested using α1(IV)NC1 in a mice model.
  
  
  RESULTS
  Like α1(IV)NC1, its N- and C- terminal α1S2(IV)NC1 and α1S1(IV)NC1 domains posses anti-proliferative, pro-apoptotic activity and inhibit ECs migration and tube formation in-vitro. Both α1S1(IV)NC1 and α1S2(IV)NC1 domains promote apoptosis by activating FasL and down stream apoptotic events including activation of caspase-8, -3 and PARP cleavage in a dose dependent manner in-vitro in ECs. Tumors in mice showed apoptotic TUNEL positive microvasculature upon α1(IV)NC1 treatment, indicating inhibition of tumor angiogenesis and tumor growth. Further, the antitumor activity of α1(IV)NC1 was abrogated when caspase-3 inhibitor was used. These results conform additional properties of α1(IV)NC1 as an endogenous angioinhibitor that induces apoptosis in-vitro and in-vivo by activating FasL mediated caspase-3.
  
  
  CONCLUSIONS
  α1(IV)NC1 and its N- and C- terminal α1S1(IV)NC1 and α1S2(IV)NC1 domains also posses pro-apoptotic and angioinhibitory activity in-vitro and in-vivo. α1(IV)NC1 regulates tumor angiogenesis by activating FasL mediated apoptosis in-vitro and in-vivo. These results demonstrate that α1(IV)NC1 and its peptides inhibit neo-vascular diseases.
authors: Irma Lerma Rangel College of Pharmacy, Texas A&M Health Science Center, Kingsville, Texas, United States of America.
authors: Raj K
authors: RK
authors: Verma
authors: Venugopal
authors: V
authors: Gunda
authors: Smita C
authors: SC
authors: Pawar
authors: Yakkanti Akul
authors: YA
authors: Sudhakar
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D053181:Caspase 8 / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D024141:Collagen Type IV / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D053222:Fas Ligand Protein / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D051379:Mice; D009389:Neovascularization, Pathologic; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism; D020033:Protein Isoforms / Q000494:pharmacology; D015398:Signal Transduction; D012878:Skin Neoplasms / Q000098:blood supply* / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D018243:Teratocarcinoma / Q000098:blood supply* / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D019014:fas Receptor / Q000235:genetics / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; D024141:Collagen Type IV; C491901:FAS protein, human; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D020033:Protein Isoforms; D019014:fas Receptor; D011065:Poly(ADP-ribose) Polymerases; D053148:Caspase 3; D053181:Caspase 8