title: VEGF antisense therapy inhibits tumor growth and improves survival in experimental pancreatic cancer.
abstract: BACKGROUND
  Vascular endothelial growth factor (VEGF), a key mediator of angiogenesis, is overexpressed in pancreatic cancer. This study evaluated VEGF production in pancreatic cancer cells and the effect of VEGF antisense on growth and angiogenesis of human pancreatic cancer in a nude mouse model.
  
  
  METHODS
  In vitro: VEGF in cell culture supernatant of pancreatic cancer cells (AsPC-1, poorly differentiated; HPAF-2, moderately differentiated) was assessed by enzyme-linked immunosorbent assay. In vivo: A VEGF antisense oligonucleotide (AS-3) was synthesized. One-mm(3) fragments of subcutaneous pancreatic cancer donor tumors were implanted into the pancreas of nude mice also receiving AS-3 (10 mg/kg/day) or vehicle intraperitoneally for 14 weeks. Primary tumor volume, metastasis, and VEGF in plasma and ascites were determined at autopsy. Microvessel density was analyzed in CD31-stained tumors.
  
  
  RESULTS
  In vitro: Both pancreatic cancer cell lines secreted VEGF protein (AsPC-1, 4200 +/- 40 pg/10(6) cells; HPAF-2, 8120 +/- 60 pg/10(6) cells). In vivo: AS-3 reduced tumor volume in the HPAF-2 group (860 +/- 140 vs 3830 +/- 590 mm(3)) and metastatic spread in both groups (AsPC-1, 6.5 +/- 0.8 vs 16.7 +/- 0.9 points; HPAF-2, 2.5 +/- 0.2 vs 8.3 +/- 1.5 points). Tumor volume was not different in the AsPC-1 group (1050 +/- 80 vs 1400 +/- 150 mm(3)). Survival was increased in the AsPC-1 group. Plasma levels of VEGF and microvessel density in tumors were significantly reduced in treated animals. Only control animals (50%) developed ascites with high VEGF concentrations.
  
  
  CONCLUSIONS
  Human pancreatic cancer cells secrete VEGF at biologically relevant high levels. AS-3 therapy normalizes plasma VEGF and decreases neoangiogenesis, thereby reducing tumor growth and metastasis and improving survival. AS-3-treated animals developed no ascites, suggesting decreased vascular permeability by reducing VEGF expression in pancreatic cancer cells.
authors.affiliation: Department of Surgery, School of Medicine, University of California-Los Angeles, Los Angeles, CA, USA. hhotz@zedat.fu-berlin.de
authors.forename: Hubert G
authors.initials: HG
authors.lastname: Hotz
authors.forename: O Joe
authors.initials: OJ
authors.lastname: Hines
authors.forename: Rizwan
authors.initials: R
authors.lastname: Masood
authors.forename: Birgit
authors.initials: B
authors.lastname: Hotz
authors.forename: Thomas
authors.initials: T
authors.lastname: Foitzik
authors.forename: Heinz J
authors.initials: HJ
authors.lastname: Buhr
authors.forename: Parkash S
authors.initials: PS
authors.lastname: Gill
authors.forename: Howard A
authors.initials: HA
authors.lastname: Reber
mesh_terms: D000818:Animals; D001201:Ascites / Q000378:metabolism; D001483:Base Sequence; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008833:Microcirculation / Q000473:pathology; D009368:Neoplasm Transplantation; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics / Q000627:therapeutic use*; D010190:Pancreatic Neoplasms / Q000098:blood supply / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D014183:Transplantation, Heterologous; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
chemical_list: D020319:Oligodeoxyribonucleotides, Antisense; D042461:Vascular Endothelial Growth Factor A