title: Phaleria macrocarpa (Boerl.) fruit induce G0/G1and G2/M cell cycle arrest and apoptosis through mitochondria-mediated pathway in MDA-MB-231 human breast cancer cell.
abstract: BACKGROUND
  Phaleria macrocarpa (Scheff) Boerl, is a well-known folk medicinal plant in Indonesia. Traditionally, P. macrocarpa has been used to control cancer, impotency, hemorrhoids, diabetes mellitus, allergies, liver and hearth disease, kidney disorders, blood diseases, acne, stroke, migraine, and various skin diseases.
  
  
  OBJECTIVE
  The purpose of this study was to determine the in situ cytotoxicity effect P. macrocarpa fruit ethyl acetate fraction (PMEAF) and the underlying molecular mechanism of cell death.
  
  
  METHODS
  MDA-MB-231 cells were incubated with PMEAF for 24h. Cell cycle and viability were examined using flow cytometry analysis. Apoptosis was determined using the Annexin V assay and also by fluorescence microscopy. Apoptosis protein profiling was detected by RayBio® Human Apoptosis Array.
  
  
  RESULTS
  The AO/PI staining and flow cytometric analysis of MDA-MB-231 cells treated with PMEAF were showed apoptotic cell death. The cell cycle analysis by flow cytometry analysis revealed that the accumulation of PMEAF treated MDA-MB-231 cells in G0/G1and G2/M-phase of the cell cycle. Moreover, the PMEAF exert cytotoxicity by increased the ROS production in MDA-MB-231 cells consistently stimulated the loss of mitochondrial membrane potential (∆Ψm) and induced apoptosis cell death by activation of numerous signalling proteins. The results from apoptosis protein profiling array evidenced that PMEAF stimulated the expression of 9 pro-apoptotic proteins (Bax, Bid, caspase 3, caspase 8, cytochrome c, p21, p27, p53 and SMAC) and suppressed the 4 anti-apoptotic proteins (Bcl-2, Bcl-w, XIAP and survivin) in MDA-MB-231 cells.
  
  
  CONCLUSIONS
  The results indicated that PMEAF treatment induced apoptosis in MDA-MB-231 cells through intrinsic mitochondrial related pathway with the participation of pro and anti-apoptotic proteins, caspases, G0/G1and G2/M-phases cell cycle arrest by p53-mediated mechanism.
authors.affiliation: Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, USM, 11800 Pulau Pinang, Malaysia.
authors.forename: Nowroji
authors.initials: N
authors.lastname: Kavitha
authors.affiliation: Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, USM, 11800 Pulau Pinang, Malaysia.
authors.forename: Chern
authors.initials: C
authors.lastname: Ein Oon
authors.affiliation: Dental Research & Training Unit, and Oral Cancer Research and Coordinating Centre (OCRCC), Faculty of Dentistry, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Yeng
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Nanomedicine-Laboratory of Immunology and Molecular Biomedical Research (LIMBR), School of Medicine (SoM), Faculty of Health, Institute for Frontier Materials (IFM), Deakin University, Waurn Ponds, VIC 3217 Australia.
authors.forename: Jagat R
authors.initials: JR
authors.lastname: Kanwar
authors.affiliation: Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, USM, 11800 Pulau Pinang, Malaysia. Electronic address: srisasidharan@yahoo.com.
authors.forename: Sreenivasan
authors.initials: S
authors.lastname: Sasidharan
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000032:analysis / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001943:Breast Neoplasms / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D005638:Fruit; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology; D010636:Phenols / Q000032:analysis; D010936:Plant Extracts / Q000032:analysis / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D029645:Thymelaeaceae
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D051017:Apoptosis Regulatory Proteins; D010636:Phenols; D010936:Plant Extracts; D017382:Reactive Oxygen Species
keywords: Apoptosis; Caspase, p53, cytochrome c; Cells cycle arrest