title: Bufalin induces mitochondrial pathway-mediated apoptosis in lung adenocarcinoma cells.
abstract: BACKGROUND
  To evaluate the effects of bufalin in A549 human lung adenocarcinoma epithelial cells in vitro and assess the underlying mechanisms.
  
  
  METHODS
  Human A549 non-small cell lung cancer (NSCLC) cells were treated with various concentrations of bufalin. Cell proliferation was measured by CCK-8 assay, apoptotic cell percentage was calculated by flow cytometry and morphological change was observed by inverted phase contrast microscopy/transmission electron microscopy. In addition, the membrane potential of mitochondria was detected by JC-1 fluorescence microscopy assay, and the related protein expression of cytochrome C and caspase-3 was analyzed by Western blotting.
  
  
  RESULTS
  Bufalin could inhibit the proliferation of A549 cells via induction of apoptosis, with the evidence of characteristic morphological changes in the nucleus and mitochondria. Furthermore, bufalin decreased the mitochondrial membrane potential with up-regulation of cytochrome C in the cytosol, and activation of caspase-3.
  
  
  CONCLUSIONS
  Bufalin inhibits the proliferation of A549 cells and triggers mitochondria-dependent apoptosis, pointing to therapeutic application for NSCLC.
authors.affiliation: The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, China E-mail : zyhong311@126.com.
authors.forename: Da-Wei
authors.initials: DW
authors.lastname: Ding
authors.forename: Yong-Hong
authors.initials: YH
authors.lastname: Zhang
authors.forename: Xin-En
authors.initials: XE
authors.lastname: Huang
authors.forename: Qing
authors.initials: Q
authors.lastname: An
authors.forename: Xun
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002018:Bufanolides / Q000494:pharmacology*; D053148:Caspase 3 / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D045304:Cytochromes c / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D015854:Up-Regulation
chemical_list: D000970:Antineoplastic Agents; D002018:Bufanolides; D045304:Cytochromes c; C505407:CASP3 protein, human; D053148:Caspase 3; C022777:bufalin