title: Plumbagin from Plumbago Zeylanica L induces apoptosis in human non-small cell lung cancer cell lines through NF- κB inactivation.
abstract: OBJECTIVE
  To detect effects of plumbagin on proliferation and apoptosis in non-small cell lung cancer cell lines, and investigate the underlying mechanisms.
  
  
  METHODS
  Human non-small cell lung cancer cell lines A549, H292 and H460 were treated with various concentrations of plumbagin. Cell proliferation rates was determined using both cell counting kit-8 (CCK-8) and clonogenic assays. Apoptosis was detected by annexin V/propidium iodide double-labeled flow cytometry and TUNEL assay. The levels of reactive oxygen species (ROS) were detected by flow cytometry. Activity of NF-κB was examined by electrophoretic mobility shift assay (EMSA) and luciferase reporter assay. Western blotting was used to assess the expression of both NF-κB regulated apoptotic-related gene and activation of p65 and IκBκ.
  
  
  RESULTS
  Plumbagin dose-dependently inhibited proliferation of the lung cancer cells. The IC50 values of plumbagin in A549, H292, and H460 cells were 10.3 μmol/L, 7.3 μmol/L, and 6.1 μmol/L for 12 hours, respectively. The compound concentration-dependently induced apoptosis of the three cell lines. Treatment with plumbagin increased the intracellular level of ROS, and inhibited the activation of NK-κB. In addition to inhibition of NF-κB/p65 nuclear translocation, the compound also suppressed the degradation of IκBκ. ROS scavenger NAC highly reversed the effect of plumbagin on apoptosis and inactivation of NK-κB in H460 cell line. Treatment with plumbagin also increased the activity of caspase-9 and caspase-3, downregulated the expression of Bcl-2, upregulated the expression of Bax, Bak, and CytC.
  
  
  CONCLUSIONS
  Plumbagin inhibits cell growth and induces apoptosis in human lung cancer cells through an NF-κB-regulated mitochondrial-mediated pathway, involving activation of ROS.
authors.affiliation: Department of Oncology, the First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
authors.forename: Tong-Peng
authors.initials: TP
authors.lastname: Xu
authors.forename: Hua
authors.initials: H
authors.lastname: Shen
authors.forename: Ling-Xiang
authors.initials: LX
authors.lastname: Liu
authors.forename: Yong-Qian
authors.initials: YQ
authors.lastname: Shu
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D049109:Cell Proliferation / Q000187:drug effects; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008175:Lung Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D009285:Naphthoquinones / Q000494:pharmacology*; D029606:Plumbaginaceae / Q000737:chemistry*; D021381:Protein Transport; D013481:Superoxides / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D016328:NF-kappa B; D009285:Naphthoquinones; D013481:Superoxides; D008156:Luciferases; C014758:plumbagin