title: Cytotoxic evaluation and induction of mitochondria-mediated apoptosis in human leukaemia HL-60 cells by Carissa spinarum stem isolate.
abstract: OBJECTIVE
  To evaluate Carissa spinarum stem isolate for its anti-cancer therapeutic potential.
  
  
  METHODS
  The n-butanol fraction of aqueous extract from Carissa spinarum stem was assessed for its cytotoxic and pro-apoptotic activity.
  
  
  RESULTS
  We report for the first time the anti-cancer potential of C. spinarum stem aqueous extract (CSE) and its n-butanol fraction (CSF). Both inhibited cell proliferation of various human cancer cell lines in which leukaemia HL-60 cells treated with CSF showed maximum growth inhibition having an inhibitory concentration (IC(50) ) value of 34.58±0.91 µg/ml. In addition, CSF induced concentration-dependent apoptosis in HL-60 cells as measured by various end-points (e.g. Annexin V binding, DNA laddering, apoptotic body formation and an increase in hypodiploid subG0 DNA content). Moreover, persistent levels of reactive oxygen species caused translocation of Bax to mitochondria and Bcl-2 degradation, which led to loss of mitochondrial membrane potential and release of cytochrome c to the cytosol. These events were associated with significant activation of caspase-3, caspase-6 and caspase-9 leading to poly (ADP-ribose) polymerase cleavage.
  
  
  CONCLUSIONS
  All the above parameters revealed that CSF induced apoptosis through the mitochondrial dependent pathway in HL-60 cells.
authors: Cancer Pharmacology Division, Indian Institute of Integrative Medicine, Jammu, India.
authors: Irum
authors: I
authors: Sehar
authors: Harish Chandra
authors: HC
authors: Pal
authors: Sangeeta
authors: S
authors: Shukla
authors: Shashi
authors: S
authors: Bhushan
authors: Abid
authors: A
authors: Hamid
authors: Bishan Dutt
authors: BD
authors: Gupta
authors: Ajit Kumar
authors: AK
authors: Saxena
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology / Q000627:therapeutic use*; D029062:Apocynaceae; D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D018922:HL-60 Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D015473:Leukemia, Promyelocytic, Acute / Q000188:drug therapy* / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology / Q000627:therapeutic use*; D018547:Plant Stems; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D010936:Plant Extracts; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D045304:Cytochromes c; D011065:Poly(ADP-ribose) Polymerases; D020169:Caspases