title: Glutamine depletion by crisantaspase hinders the growth of human hepatocellular carcinoma xenografts.
abstract: BACKGROUND
  A subset of human hepatocellular carcinomas (HCC) exhibit mutations of β-catenin gene CTNNB1 and overexpress Glutamine synthetase (GS). The CTNNB1-mutated HCC cell line HepG2 is sensitive to glutamine starvation induced in vitro with the antileukemic drug Crisantaspase and the GS inhibitor methionine-L-sulfoximine (MSO).
  
  
  METHODS
  Immunodeficient mice with subcutaneous xenografts of the CTNNB1-mutated HCC cell lines HepG2 and HC-AFW1 were treated with Crisantaspase and/or MSO, and tumour growth was monitored. At the end of treatment, tumour weight and histology were assessed. Serum and tissue amino acids were determined by HPLC. Gene and protein expression were estimated with RT-PCR and western blot and GS activity with a colorimetric method. mTOR activity was evaluated from the phosphorylation of p70S6K1.
  
  
  RESULTS
  Crisantaspase and MSO depleted serum glutamine, lowered glutamine in liver and tumour tissue, and inhibited liver GS activity. HepG2 tumour growth was significantly reduced by either Crisantaspase or MSO, and completely suppressed by the combined treatment. The combined treatment was also effective against xenografts of the HC-AFW1 cell line, which is Crisantaspase resistant in vitro.
  
  
  CONCLUSIONS
  The combination of Crisantaspase and MSO reduces glutamine supply to CTNNB1-mutated HCC xenografts and hinders their growth.
authors: Department of Biomedical Biotechnological and Translational Sciences (SBiBiT), University of Parma, Parma, Italy.
authors: M
authors: M
authors: Chiu
authors: Department of Biomedical Biotechnological and Translational Sciences (SBiBiT), University of Parma, Parma, Italy.
authors: S
authors: S
authors: Tardito
authors: Department of Experimental and Clinical Medicine, University of Firenze, Firenze, Italy.
authors: S
authors: S
authors: Pillozzi
authors: Department of Experimental and Clinical Medicine, University of Firenze, Firenze, Italy.
authors: A
authors: A
authors: Arcangeli
authors: Department of Biomedical Biotechnological and Translational Sciences (SBiBiT), University of Parma, Parma, Italy.
authors: A
authors: A
authors: Armento
authors: Department of Biomedical Biotechnological and Translational Sciences (SBiBiT), University of Parma, Parma, Italy.
authors: J
authors: J
authors: Uggeri
authors: Unit of Infectious Diseases and Hepatology, Azienda Ospedaliero-Universitaria di Parma, Parma, Italy.
authors: G
authors: G
authors: Missale
authors: Department of Biomedical Biotechnological and Translational Sciences (SBiBiT), University of Parma, Parma, Italy.
authors: M G
authors: MG
authors: Bianchi
authors: Department of Biomedical Biotechnological and Translational Sciences (SBiBiT), University of Parma, Parma, Italy.
authors: A
authors: A
authors: Barilli
authors: Department of Biomedical Biotechnological and Translational Sciences (SBiBiT), University of Parma, Parma, Italy.
authors: V
authors: V
authors: Dall'Asta
authors: Department of Biomedical Biotechnological and Translational Sciences (SBiBiT), University of Parma, Parma, Italy.
authors: N
authors: N
authors: Campanini
authors: 1] Department of Biomedical Biotechnological and Translational Sciences (SBiBiT), University of Parma, Parma, Italy [2] Center for Translational and Molecular Oncology (COMT), University of Parma, Parma, Italy.
authors: E M
authors: EM
authors: Silini
authors: Department of Pediatric Surgery and Urology, University Childreńs Hospital, Eberhard Karls University Tübingen, Tübingen, Germany.
authors: J
authors: J
authors: Fuchs
authors: Department of Pediatric Surgery and Urology, University Childreńs Hospital, Eberhard Karls University Tübingen, Tübingen, Germany.
authors: S
authors: S
authors: Armeanu-Ebinger
authors: Department of Biomedical Biotechnological and Translational Sciences (SBiBiT), University of Parma, Parma, Italy.
authors: O
authors: O
authors: Bussolati
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000627:therapeutic use; D001215:Asparaginase / Q000494:pharmacology* / Q000627:therapeutic use*; D001216:Asparagine / Q000097:blood; D015820:Cadherins / Q000032:analysis; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D004791:Enzyme Inhibitors / Q000627:therapeutic use; D005974:Glutamate-Ammonia Ligase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D005973:Glutamine / Q000032:analysis / Q000097:blood; D056945:Hep G2 Cells; D006801:Humans; D019394:Ki-67 Antigen / Q000032:analysis; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D008297:Male; D008717:Methionine Sulfoximine / Q000627:therapeutic use; D051379:Mice; D008819:Mice, Nude; D009154:Mutation; D047368:Tumor Burden / Q000187:drug effects*; D023041:Xenograft Model Antitumor Assays; D051176:beta Catenin / Q000032:analysis / Q000235:genetics*
chemical_list: D000970:Antineoplastic Agents; C495270:CTNNB1 protein, human; D015820:Cadherins; D004791:Enzyme Inhibitors; D019394:Ki-67 Antigen; D051176:beta Catenin; D005973:Glutamine; D008717:Methionine Sulfoximine; D001216:Asparagine; D001215:Asparaginase; D005974:Glutamate-Ammonia Ligase