title: In vitro sensitivity of human gastric cancer cells (HGC-27) to Helicobacter pylori cytotoxin.
abstract: BACKGROUND
  The VacA cytotoxin produced by Helicobacter pylori is considered an important co-factor in the pathogenesis of chronic gastritis, peptic ulcer and gastric carcinoma. The toxin remains partly bound on the bacterial surface, but a certain amount is secreted and can bind receptors on gastric epithelium. The vacuolizing activity of this toxin is related to alteration of endo-lysosomic function and pore formation into plasmatic membrane.
  
  
  METHODS
  We investigated the 'in vitro' effect of filtrates obtained from two broth cultures of H. pylori with different genotype (vacA+ and vacA-) as verified by PCR. The effect was studied on three cell lines of epithelial origin: HeLa cells (reference strain for testing vacuolization), human transformed keratinocytes HaCaT, human gastric carcinoma cells HGC-27, and on a murine leukaemia WEHI-3B. The filtrate concentrations capable of giving vacuolization (NRU test), antiproliferative and cytotoxic effects (MTT test) were determined. The modulating effect of filtrates on drug toxicity was investigated on HeLa and HGC-27 cells by testing topoisomerase inhibitors (Ciprofloxacin and Camptothecin) and non-steroidal anti-inflammatory molecules (Aspirin and Indomethacin).
  
  
  RESULTS
  Our results confirm that vacuolizing activity is present only in VacA+ filtrate and that HaCaT and HeLa cells show a similar sensitivity, whereas gastric HGC-27 cells appear significantly resistant to VacA+ activity. Although VacA filtrate does not produce vacuolisation, it affects the cell proliferation and is cytotoxic to the four cell lines. Both the VacA+ and VacA- filtrates (at non-cytotoxic concentrations) produce a decrease in drug toxicity with the unique exception of Ciprofloxacin to gastric HGC-27 cells, which in the presence of VacA+ and VacA- produces a significant increase in toxicity.
  
  
  CONCLUSIONS
  These data suggest that products from H. pylori (other than those that have antiproliferative and toxic activity) may modulate the sensitivity of cells to drugs 'in vitro'. If this also occurs 'in vivo', we can assume that H. pylori products interfere with drug activity on gastric tissue and also with other factors (such as cytokines) with a role in the genesis of diseases in which Helicobacters are potentially involved.
authors.affiliation: Institute of Microbiology, University of Milan, Italy. augusto.pessina@unimi.it
authors.forename: A
authors.initials: A
authors.lastname: Pessina
authors.forename: M
authors.initials: M
authors.lastname: Bayo
authors.forename: C
authors.initials: C
authors.lastname: Croera
authors.forename: F
authors.initials: F
authors.lastname: Meringolo
authors.forename: M G
authors.initials: MG
authors.lastname: Neri
authors.forename: L
authors.initials: L
authors.lastname: Montesissa
authors.forename: A
authors.initials: A
authors.lastname: Raimondi
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000633:toxicity; D001426:Bacterial Proteins / Q000494:pharmacology*; D001427:Bacterial Toxins / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003114:Colony-Forming Units Assay; D003603:Cytotoxins / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D006367:HeLa Cells; D016480:Helicobacter pylori; D006801:Humans; D013274:Stomach Neoplasms / Q000473:pathology / Q000648:ultrastructure*; D059004:Topoisomerase I Inhibitors; D014617:Vacuoles / Q000187:drug effects*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D003603:Cytotoxins; D059004:Topoisomerase I Inhibitors; C074689:VacA protein, Helicobacter pylori