title: Chloroquine-mediated cell death in metastatic pancreatic adenocarcinoma through inhibition of autophagy.
abstract: BACKGROUND
  Cells in the interior of pancreatic tumors are believed to undergo continual autophagy to maintain homeostasis during unregulated growth in hypoxia caused by impaired vascularity. We hypothesize that treating metastatic cells with chloroquine, an inhibitor of autophagy, in hypoxia will decrease cell viability and induce cell death.
  
  
  METHODS
  MiaPaCa2 (non-metastatic) and S2VP10 (metastatic) cell lines were treated with 25 and 50 µM chloroquine for 24 and 48 hours in normoxia and hypoxia (5-10% O₂). Viability was measured using ATPlite™. After treatment, the cell stress was analyzed, and protein was lysed and quantified using the Bradford assay. Autophagy-associated protein levels were determined by Western blot.
  
  
  RESULTS
  S2VP10 cells treated for 48 hours with 50 μM chloroquine in hypoxia had 24% viability compared to normoxia control, with loss of 10% viability caused by low O₂ alone. MiaPaCa2 cells under these conditions had 60% viability compared to normoxia control, with loss of 25% viability caused by low O₂ alone. Analysis of cell stress pathways and dosimetry of Western blot data suggest that chloroquine inhibits the autophagy pathway in the metastatic S2VP10 cells.
  
  
  CONCLUSIONS
  Autophagy blockage with chloroquine or similar-acting drugs may serve as a viable therapy for highly metastatic pancreatic cancers.
authors.affiliation: Department of Bioengineering, University of Louisville. Louisville, KY, USA. lrmcna01@louisville.edu.
authors.forename: Hermann B
authors.initials: HB
authors.lastname: Frieboes
authors.forename: Justin S
authors.initials: JS
authors.lastname: Huang
authors.forename: Wenyuan C
authors.initials: WC
authors.lastname: Yin
authors.forename: Lacey R
authors.initials: LR
authors.lastname: McNally
mesh_terms: D000230:Adenocarcinoma / Q000473:pathology* / Q000503:physiopathology; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001343:Autophagy / Q000187:drug effects* / Q000502:physiology; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002738:Chloroquine / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D006706:Homeostasis / Q000187:drug effects / Q000502:physiology; D006801:Humans; D000860:Hypoxia / Q000503:physiopathology; D009362:Neoplasm Metastasis / Q000473:pathology* / Q000503:physiopathology; D010190:Pancreatic Neoplasms / Q000473:pathology* / Q000503:physiopathology; D013997:Time Factors
chemical_list: D002738:Chloroquine