title: Harmine activates intrinsic and extrinsic pathways of apoptosis in B16F-10 melanoma.
abstract: BACKGROUND
  Harmine is a beta-carboline alkaloid from the plant Peganum harmala. Previous studies found that harmine inhibited metastasis of B16F-10 melanoma cells. This study aims to elucidate the role of harmine in apoptosis of B16F-10 cells.
  
  
  METHODS
  B16F-10 melanoma cells were treated in the presence and absence of harmine in vitro. Morphological changes, cell cycle and expression of various pro and anti- apoptotic genes were analyzed for the study of apoptosis.
  
  
  RESULTS
  Morphological observation and DNA laddering assay showed that harmine treated cells displayed marked apoptotic characteristics, such as nuclear fragmentation, appearance of apoptotic bodies and DNA laddering fragment. TUNEL assay and flow cytometric analysis also confirmed apoptosis. Furthermore, RT-PCR analysis showed that harmine induced apoptosis in B16F-10 melanoma cells by up-regulating Bax and activating Caspase-3, 9 and p53 and down-regulating Bcl-2. Harmine also up-regulated Caspase-8 and Bid, indicating that harmine affected both extrinsic and intrinsic pathways of apoptosis. This study also showed inhibitory effects of harmine on some transcription factors and pro- inflammatory cytokines that protect cell from apoptosis.
  
  
  CONCLUSIONS
  Harmine activates both intrinsic and extrinsic pathways of apoptosis and regulates some transcription factors and pro-inflammatory cytokines.
authors.affiliation: Amala Cancer Research Centre, Amala Nagar, Thrissur, Kerala, India, 680555. girijakuttan@gmail.com.
authors.forename: Thayele Purayil
authors.initials: TP
authors.lastname: Hamsa
authors.forename: Girija
authors.initials: G
authors.lastname: Kuttan