title: Arginine-enhanced enteral nutrition augments the growth of a nitric oxide-producing tumor.
abstract: BACKGROUND
  Arginine-enhanced diets have been shown to be beneficial in tumor-bearing hosts, but no data exist regarding their effects in hosts bearing nitric oxide (NO)-producting tumors.
  
  
  OBJECTIVE
  To examine the effect of arginine supplementation on the growth of a NO-producing murine breast cancer cell line.
  
  
  METHODS
  EMT-6 cells were grown in various concentrations of arginine in the presence or absence of the inducible nitric oxide synthase (iNOS) inhibitor, aminoguanidine (1 mmol/L). Forty-eight hours later, nitrite accumulation and viable cell number were assessed. BALB/c mice were then pair-fed basal purified diets (n = 10), 4% casein diets (isonitrogenous control, n = 5), or 4% arginine-enhanced diets (n = 10). One week later, 10(5) EMT-6 cells were implanted subcutaneously into the dorsal flank. After tumor implantation, five mice fed basal purified diets and five mice fed arginine-enhanced diets also received aminoguanidine (100 mg/kg subcutaneously twice daily). Two weeks after tumor cell implantation, tumor size (mean diameter), animal weight, serum and tumor nitrite and nitrate levels were measured.
  
  
  RESULTS
  There was minimal nitrite accumulation in arginine-free media, while increasing the arginine concentration increased nitrite levels. Viable cell number did not increase in arginine-free media, but increased nearly twofold in 100 and 1000 mumol/L arginine. In 5000 and 10,000 mumol/L arginine, the difference in viable cell number was not statistically different than that seen in arginine-free media, whereas the addition of aminoguanidine blocked nitrite accumulation and increased viable cell number at these arginine concentrations. Arginine-enhanced diets stimulated tumor growth in vivo more than twofold over tumor growth in mice fed isonitrogenous control or basal purified enteral diets. Mice fed arginine-enhanced diets also had increased serum nitrite and nitrate levels over mice fed basal purified enteral diets, whereas tumors from mice fed arginine-enhanced diets had nitrite and nitrate levels similar to mice fed basal purified enteral diets. Aminoguanidine blocked the increase in serum nitrite and nitrate, but failed to block the increased tumor growth in mice receiving the arginine-supplemented diets.
  
  
  CONCLUSIONS
  Arginine concentration influences the growth of EMT-6 tumor cells in vitro and dietary arginine supplementation augments tumor growth in vivo. The mechanism of the growth modulation in vitro is NO-dependent whereas the enhanced tumor growth in vivo is NO-independent.
authors.affiliation: Department of Surgery, University of Florida College of Medicine, Gainesville 32610-00286, USA.
authors.forename: P D
authors.initials: PD
authors.lastname: Edwards
authors.forename: D
authors.initials: D
authors.lastname: Topping
authors.forename: M I
authors.initials: MI
authors.lastname: Kontaridis
authors.forename: L L
authors.initials: LL
authors.lastname: Moldawer
authors.forename: E M
authors.initials: EM
authors.lastname: Copeland
authors.forename: D S
authors.initials: DS
authors.lastname: Lind
mesh_terms: D000818:Animals; D001120:Arginine / Q000008:administration & dosage*; D002455:Cell Division; D004750:Enteral Nutrition; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006146:Guanidines / Q000494:pharmacology; D008325:Mammary Neoplasms, Experimental / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009368:Neoplasm Transplantation; D009566:Nitrates / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D009573:Nitrites / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D004791:Enzyme Inhibitors; D006146:Guanidines; D009566:Nitrates; D009573:Nitrites; D009569:Nitric Oxide; D001120:Arginine; D019001:Nitric Oxide Synthase; C004479:pimagedine