title: Differential metabolomic analysis of the potential antiproliferative mechanism of olive leaf extract on the JIMT-1 breast cancer cell line.
abstract: A new differential metabolomic approach has been developed to identify the phenolic cellular metabolites derived from breast cancer cells treated with a supercritical fluid extracted (SFE) olive leaf extract. The SFE extract was previously shown to have significant antiproliferative activity relative to several other olive leaf extracts examined in the same model. Upon SFE extract incubation of JIMT-1 human breast cancer cells, major metabolites were identified by using HPLC coupled to electrospray ionization quadrupole-time-of-flight mass spectrometry (ESI-Q-TOF-MS). After treatment, diosmetin was the most abundant intracellular metabolite, and it was accompanied by minor quantities of apigenin and luteolin. To identify the putative antiproliferative mechanism, the major metabolites and the complete extract were assayed for cell cycle, MAPK and PI3K proliferation pathways modulation. Incubation with only luteolin showed a significant effect in cell survival. Luteolin induced apoptosis, whereas the whole olive leaf extract incubation led to a significant cell cycle arrest at the G1 phase. The antiproliferative activity of both pure luteolin and olive leaf extract was mediated by the inactivation of the MAPK-proliferation pathway at the extracellular signal-related kinase (ERK1/2). However, the flavone concentration of the olive leaf extract did not fully explain the strong antiproliferative activity of the extract. Therefore, the effects of other compounds in the extract, probably at the membrane level, must be considered. The potential synergistic effects of the extract also deserve further attention. Our differential metabolomics approach identified the putative intracellular metabolites from a botanical extract that have antiproliferative effects, and this metabolomics approach can be expanded to other herbal extracts or pharmacological complex mixtures.
authors.affiliation: Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, Avda. Universidad s/n, 03202 Elche, Spain.
authors.forename: Enrique
authors.initials: E
authors.lastname: Barrajón-Catalán
authors.affiliation: Laboratoire de Biotechnologie de l'Olivier, Centre de Biotechnologie de Borj Cedria, BP 901, 2050 Hammam-Lif, Tunisia.
authors.forename: Amani
authors.initials: A
authors.lastname: Taamalli
authors.affiliation: Department of Analytical Chemistry, Faculty of Sciences, University of Granada, Granada 18071, Spain; Research and Development of Functional Food Centre (CIDAF), Health Science Technological Park, Avda. del Conocimiento s/n, 18100 Granada, Spain.
authors.forename: Rosa
authors.initials: R
authors.lastname: Quirantes-Piné
authors.affiliation: Department of Analytical Chemistry, Faculty of Sciences, University of Granada, Granada 18071, Spain; Research and Development of Functional Food Centre (CIDAF), Health Science Technological Park, Avda. del Conocimiento s/n, 18100 Granada, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Roldan-Segura
authors.affiliation: Department of Analytical Chemistry, Faculty of Sciences, University of Granada, Granada 18071, Spain; Research and Development of Functional Food Centre (CIDAF), Health Science Technological Park, Avda. del Conocimiento s/n, 18100 Granada, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Arráez-Román
authors.affiliation: Department of Analytical Chemistry, Faculty of Sciences, University of Granada, Granada 18071, Spain; Research and Development of Functional Food Centre (CIDAF), Health Science Technological Park, Avda. del Conocimiento s/n, 18100 Granada, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Segura-Carretero
authors.affiliation: Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, Avda. Universidad s/n, 03202 Elche, Spain. Electronic address: vmicol@umh.es.
authors.forename: Vicente
authors.initials: V
authors.lastname: Micol
authors.affiliation: Laboratoire de Biotechnologie de l'Olivier, Centre de Biotechnologie de Borj Cedria, BP 901, 2050 Hammam-Lif, Tunisia.
authors.forename: Mokhtar
authors.initials: M
authors.lastname: Zarrouk
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects; D055432:Metabolomics; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D031658:Olea / Q000737:chemistry*; D010936:Plant Extracts / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology*; D018515:Plant Leaves / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D010936:Plant Extracts; D020928:Mitogen-Activated Protein Kinases
keywords: Diosmetin; HPLC–ESI-Q-TOF-MS; JIMT-1 breast cancer cells; Luteolin; MAPK; Metabolomics