title: Increased expression of neprilysin (neutral endopeptidase 24.11) in rat and human hepatocellular carcinomas.
abstract: BACKGROUND
  Neprilysin (EC 3.4.24.11) (NEP), a membrane metallopeptidase, is identical with common acute lymphoblastic leukemia antigen or cluster differentiation antigen 10. This antigen is present in blast cells in acute lymphoblastic leukemias and is implicated in differentiation of B lymphocytes. NEP cleaves a variety of peptides including bradykinin, substance P, bombesin, enkephalins, and atrial natriuretic peptide. We investigated its expression in several variants of rat hepatomas and a human hepatocellular carcinoma cell line. Normal rat and human livers were used as controls.
  
  
  METHODS
  The expression of NEP (common acute lymphoblastic leukemia antigen) was determined with: (a) enzyme assays; (b) high performance liquid chromatography analysis of bradykinin metabolism; (c) immunoprecipitation; and (d) mRNA characterization.
  
  
  RESULTS
  NEP activity increased by 2 to 3 orders of magnitude in all rat hepatomas and in the human SK-HEP1 cell line, compared with normal tissues. Antiserum against rat NEP precipitated 93% of endopeptidase activity in rat hepatomas, whereas monoclonal antibody to common acute lymphoblastic leukemia antigen immunoprecipitated 99% of that in human hepatocarcinoma cells. Solubilized rat hepatoma membranes cleaved bradykinin to a hepta- and dipeptide; the reaction was inhibited by an NEP inhibitor. Activity of three other membrane peptidases did not increase in rat hepatomas. Northern hybridization revealed the presence of NEP mRNA in rat hepatoma, but not in normal liver. Reverse transcriptase-polymerase chain reaction showed that hepatomas have higher amounts of NEP mRNA than normal liver of the same strain.
  
  
  CONCLUSIONS
  Rat hepatomas and a human hepatocarcinoma cell line express high amounts of NEP, in contrast to normal rat and human livers, which have very little. The increase in NEP activity could be due to increased transcription by tumor cells and may signal malignant transformation of liver cells.
authors.affiliation: Laboratory of Peptide Research/Department of Pharmacology, University of Illinois College of Medicine, Chicago.
authors.forename: T
authors.initials: T
authors.lastname: Dragović
authors.forename: P A
authors.initials: PA
authors.lastname: Deddish
authors.forename: F
authors.initials: F
authors.lastname: Tan
authors.forename: G
authors.initials: G
authors.lastname: Weber
authors.forename: E G
authors.initials: EG
authors.lastname: Erdös
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D001920:Bradykinin / Q000032:analysis / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000737:chemistry / Q000201:enzymology* / Q000473:pathology; D002462:Cell Membrane / Q000737:chemistry / Q000201:enzymology / Q000648:ultrastructure; D002851:Chromatography, High Pressure Liquid; D015342:DNA Probes; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000201:enzymology; D008113:Liver Neoplasms / Q000737:chemistry / Q000201:enzymology* / Q000473:pathology; D008114:Liver Neoplasms, Experimental / Q000737:chemistry / Q000201:enzymology* / Q000473:pathology; D015260:Neprilysin / Q000032:analysis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D012334:RNA, Neoplasm / Q000032:analysis / Q000235:genetics; D051381:Rats; D011912:Rats, Inbred ACI; D011915:Rats, Inbred BUF; D014407:Tumor Cells, Cultured
chemical_list: D015342:DNA Probes; D012334:RNA, Neoplasm; D015260:Neprilysin; D001920:Bradykinin