title: A Petiveria alliacea standardized fraction induces breast adenocarcinoma cell death by modulating glycolytic metabolism.
abstract: BACKGROUND
  Folk medicine uses aqueous and alcoholic extracts from Petiveria alliacea (Phytolaccaceae) in leukemia and breast cancer treatment in the Caribbean, Central and South America. Herein, we validated the biological activity of a Petiveria alliacea fraction using a metastatic breast adenocarcinoma model (4T1).
  
  
  METHODS
  Petiveria alliacea fraction biological activity was determined estimating cell proliferation, cell colony growth capacity and apoptosis (caspase-3 activity, DNA fragmentation and mitochondrial membrane potential) in 4T1 cells. Petiveria alliacea was used at IC₅₀ concentration (29 µg/mL) and 2 dilutions below, doxorubicin at 0.27 µg/mL (positive control) and dibenzyl disulfide at 2.93 µg/mL (IC50 fraction marker compound). Proteomic estimations were analyzed by LC-MS-MS. Protein level expression was confirmed by RT-PCR. Glucose and lactate levels were measured by enzymatic assays. LD50 was established in BALB/c mice and antitumoral activity evaluated in mice transplanted with GFP-tagged 4T1 cells. Mice were treated with Petiveria alliacea fraction via I.P (182 mg/kg corresponding to 1/8 of LD₅₀ and 2 dilutions below).
  
  
  RESULTS
  Petiveria alliacea fraction in vitro induces 4T1 cells apoptosis, caspase-3 activation, DNA fragmentation without mitochondria membrane depolarization, and decreases cell colony growth capacity. Also, changes in glycolytic enzymes expression cause a decrease in glucose uptake and lactate production. Fraction also promotes breast primary tumor regression in BALB/c mice transplanted with GFP-tagged 4T1 cells.
  
  
  CONCLUSIONS
  A fraction of Petiveria alliacea leaves and stems induces in vitro cell death and in vivo tumor regression in a murine breast cancer model. Our results validate in partly, the traditional use of Petiveria alliacea in breast cancer treatment, revealing a new way of envisioning Petiveria alliacea biological activity. The fraction effect on the glycolytic pathway enzymes contributes to explain the antiproliferative and antitumor activities.
authors.affiliation: Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Pontificia Universidad Javeriana, Carrera 7 N. 43-82 Building 52, Office 608, Bogotá, Colombia.
authors.forename: John Fredy
authors.initials: JF
authors.lastname: Hernández
authors.affiliation: Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Pontificia Universidad Javeriana, Carrera 7 N. 43-82 Building 52, Office 608, Bogotá, Colombia.
authors.forename: Claudia Patricia
authors.initials: CP
authors.lastname: Urueña
authors.affiliation: Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Pontificia Universidad Javeriana, Carrera 7 N. 43-82 Building 52, Office 608, Bogotá, Colombia.
authors.forename: Maria Claudia
authors.initials: MC
authors.lastname: Cifuentes
authors.affiliation: Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Pontificia Universidad Javeriana, Carrera 7 N. 43-82 Building 52, Office 608, Bogotá, Colombia.
authors.forename: Tito Alejandro
authors.initials: TA
authors.lastname: Sandoval
authors.affiliation: Grupo de Farmacología Vegetal, Fundación Universitaria Juan N. Corpas, Carrera 111N. 159A-61, Bogotá, Colombia.
authors.forename: Luis Miguel
authors.initials: LM
authors.lastname: Pombo
authors.affiliation: Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Pontificia Universidad Javeriana, Carrera 7 N. 43-82 Building 52, Office 608, Bogotá, Colombia.
authors.forename: Diana
authors.initials: D
authors.lastname: Castañeda
authors.affiliation: Department of Microbiology, Biochemistry & Immunology, Morehouse School of Medicine, 720 Westview Drive, Atlanta, GA 30310, USA.
authors.forename: Alexzander
authors.initials: A
authors.lastname: Asea
authors.affiliation: Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Pontificia Universidad Javeriana, Carrera 7 N. 43-82 Building 52, Office 608, Bogotá, Colombia. Electronic address: susana.fiorentino@javeriana.edu.co.
authors.forename: Susana
authors.initials: S
authors.lastname: Fiorentino
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy*; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use; D001943:Breast Neoplasms / Q000188:drug therapy*; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D053938:DNA Fragmentation; D005260:Female; D006019:Glycolysis / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D029602:Phytolaccaceae; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology / Q000627:therapeutic use
chemical_list: D000970:Antineoplastic Agents; D010936:Plant Extracts; D053148:Caspase 3
keywords: Breast adenocarcinoma cells 4T1; Cell death; Natural products; Petiveria alliacea (Phytolaccaceae); Warburg effect