title: The mechanisms of nadroparin-mediated inhibition of proliferation of two human lung cancer cell lines.
abstract: OBJECTIVE
  Clinical data suggest that heparin treatment improves survival of lung cancer patients, but the mechanisms involved are not fully understood. We investigated whether low molecular weight heparin nadroparin, directly affects lung cancer cell population growth in conventionally cultured cell lines.
  
  
  METHODS
  A549 and CALU1 cells' viability was assessed by MTT and trypan blue exclusion assays. Cell proliferation was assessed using 5-bromo-2-deoxyuridine incorporation. Apoptosis and cell-cycle distribution were analysed by flow cytometry; cyclin B1, Cdk1, p-Cdk1 Cdc25C, p-Cdc25C and p21 expressions were analysed by western blotting. mRNA levels were analysed by real time RT-PCR.
  
  
  RESULTS
  Nadroparin inhibited cell proliferation by 30% in both cell lines; it affected the cell cycle in A549, but not in CALU-1 cells, inducing arrest in the G(2) /M phase. Nadroparin in A549 culture inhibited cyclin B1, Cdk1, Cdc25C and p-Cdc25C, while levels of p-Cdk1 were elevated; p21 expression was not altered. Dalteparin caused a similar reduction in A549 cell population growth; however, it did not alter cyclin B1 expression as expected, based on previous reports. Fondaparinux caused minimal inhibition of A549 cell population growth and no effect on either cell cycle or cyclin B1 expression.
  
  
  CONCLUSIONS
  Nadroparin inhibited proliferation of A549 cells by inducing G(2) /M phase cell-cycle arrest that was dependent on the Cdc25C pathway, whereas CALU-1 cell proliferation was halted by as yet not elucidated modes.
authors.affiliation: Laboratory of Respiratory Cell Biology, Cardiac, Thoracic and Vascular Department, University of Pisa and University Hospital of Pisa, Pisa, Italy.
authors.forename: Y
authors.initials: Y
authors.lastname: Carmazzi
authors.forename: M
authors.initials: M
authors.lastname: Iorio
authors.forename: C
authors.initials: C
authors.lastname: Armani
authors.forename: S
authors.initials: S
authors.lastname: Cianchetti
authors.forename: F
authors.initials: F
authors.lastname: Raggi
authors.forename: T
authors.initials: T
authors.lastname: Neri
authors.forename: C
authors.initials: C
authors.lastname: Cordazzo
authors.forename: S
authors.initials: S
authors.lastname: Petrini
authors.forename: R
authors.initials: R
authors.lastname: Vanacore
authors.forename: F
authors.initials: F
authors.lastname: Bogazzi
authors.forename: P
authors.initials: P
authors.lastname: Paggiaro
authors.forename: A
authors.initials: A
authors.lastname: Celi
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy* / Q000378:metabolism; D000925:Anticoagulants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy* / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D005434:Flow Cytometry; D006801:Humans; D008168:Lung / Q000166:cytology; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D017762:Nadroparin / Q000494:pharmacology*; D020687:cdc25 Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000925:Anticoagulants; D017762:Nadroparin; C517009:CDC25C protein, human; D020687:cdc25 Phosphatases