title: Tetracycline-inducible protein expression in pancreatic cancer cells: effects of CapG overexpression.
abstract: OBJECTIVE
  To establish stable tetracycline-inducible pancreatic cancer cell lines.
  
  
  METHODS
  Suit-2, MiaPaca-2, and Panc-1 cells were transfected with a second generation reverse tetracycline-controlled transactivator protein (rtTA2S-M2), under the control of either a cytomegalovirus (CMV) or a chicken β-actin promoter, and the resulting clones were characterised.
  
  
  RESULTS
  Use of the chicken (β-actin) promoter proved superior for both the production and maintenance of doxycycline-inducible cell lines. The system proved versatile, enabling transient inducible expression of a variety of genes, including GST-P, CYP2E1, S100A6, and the actin capping protein, CapG. To determine the physiological utility of this system in pancreatic cancer cells, stable inducible CapG expressors were established. Overexpressed CapG was localised to the cytoplasm and the nuclear membrane, but was not observed in the nucleus. High CapG levels were associated with enhanced motility, but not with changes to the cell cycle, or cellular proliferation. In CapG-overexpressing cells, the levels and phosphorylation status of other actin-moduating proteins (Cofilin and Ezrin/Radixin) were not altered. However, preliminary analyses suggest that the levels of other cellular proteins, such as ornithine aminotransferase and enolase, are altered upon CapG induction.
  
  
  CONCLUSIONS
  We have generated pancreatic-cancer derived cell lines in which gene expression is fully controllable.
authors.affiliation: Liverpool CR-UK Centre, Department of Molecular and Therapeutic Cancer Medicine, University of Liverpool, Liverpool, L69 3GA, UK.
authors.forename: Sarah
authors.initials: S
authors.lastname: Tonack
authors.forename: Sabina
authors.initials: S
authors.lastname: Patel
authors.forename: Mehdi
authors.initials: M
authors.lastname: Jalali
authors.forename: Taoufik
authors.initials: T
authors.lastname: Nedjadi
authors.forename: Rosalind E
authors.initials: RE
authors.lastname: Jenkins
authors.forename: Christopher
authors.initials: C
authors.lastname: Goldring
authors.forename: John
authors.initials: J
authors.lastname: Neoptolemos
authors.forename: Eithne
authors.initials: E
authors.lastname: Costello
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002453:Cell Cycle / Q000502:physiology; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D002645:Chickens; D004318:Doxycycline / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D013752:Tetracycline / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D008840:Microfilament Proteins; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; C066575:CAPG protein, human; D013752:Tetracycline; D004318:Doxycycline