title: Marrubium vulgare ethanolic extract induces proliferation block, apoptosis, and cytoprotective autophagy in cancer cells in vitro.
abstract: Marrubium vulgare is a European medicinal plant with numerous beneficial effects on human health. The aim of the study was to isolate the plant ethanolic extract (MVE) and to investigate its anti-melanoma and anti-glioma effects. MVE was prepared by the modified pharmacopoeial percolation method and characterized by UHPLC-LTQ OrbiTrap MS. MVE dose-dependently reduced viability of melanoma (B16) and glioma (U251) cells, but not peripheral blood mononuclear cells. It arrested cell cycle in S+G2/M phase, which was associated with the activation of MAP kinase p38 and up-regulation of antiproliferative genes p53, p21 and p27. MVE induced oxidative stress, while antioxidants abrogated its antitumor effect. Furthermore, MVE induced mitochondrial depolarization, activation of caspase-9 and -3, Parp cleavage, phosphatidylserine exposure and DNA fragmentation. The mitochondrial apoptotic pathway was associated with the up-regulation of proapoptotic genes Pten, Bak1, Apaf1, and Puma and down-regulation of antiapoptotic genes survivin and Xiap. MVE also stimulated the expression of autophagy-related genes Atg5, Atg7, Atg12, Beclin-1, Gabarab and Sqstm1, as well as LC3-I conversion to the autophagosome associated LC3-II, while autophagy inhibitors exacerbated its cytotoxicity. Finally, the most abundant phenolic components of MVE, ferulic, p-hydroxybenzoic, caffeic and chlorogenic acids, did not exert a profound effect on viability of tumor cells, suggesting that other components individually or in concert are the mediators of the extracts' cytotoxicity. By demonstrating the ability of MVE to inhibit proliferation, induce apoptosis and cytoprotective autophagy, our results suggest that MVE, alone or combined with autophagy inhibitors, could be a good candidate for anti-melanoma and anti-glioma therapy.
authors.affiliation: University of Belgrade Institute of Microbiology and Immunology, School of Medicine Belgrade Serbia vericapaunovic@gmail.com.
authors.forename: V
authors.initials: V
authors.lastname: Paunovic
authors.affiliation: University of Belgrade Institute of Microbiology and Immunology, School of Medicine Belgrade Serbia.
authors.forename: M
authors.initials: M
authors.lastname: Kosic
authors.affiliation: Institute for Medicinal Plant Research Dr Josif Pancic Department for Pharmaceutical Research and Development Belgrade Serbia.
authors.forename: S
authors.initials: S
authors.lastname: Djordjevic
authors.affiliation: Institute for Medicinal Plant Research Dr Josif Pancic Department for Pharmaceutical Research and Development Belgrade Serbia.
authors.forename: A
authors.initials: A
authors.lastname: Zugic
authors.affiliation: University of Belgrade Institute of Microbiology and Immunology, School of Medicine Belgrade Serbia.
authors.forename: N
authors.initials: N
authors.lastname: Djalinac
authors.affiliation: University of Belgrade Faculty of Chemistry Belgrade Serbia.
authors.forename: U
authors.initials: U
authors.lastname: Gasic
authors.affiliation: University of Belgrade Institute of Microbiology and Immunology, School of Medicine Belgrade Serbia.
authors.forename: V
authors.initials: V
authors.lastname: Trajkovic
authors.affiliation: University of Belgrade Institute for Biological Research Sinisa Stankovic Belgrade Serbia buajk@yahoo.com.
authors.forename: J
authors.initials: J
authors.lastname: Harhaji-Trajkovic
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects*; D020169:Caspases / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D015536:Down-Regulation / Q000187:drug effects; D000431:Ethanol / Q000737:chemistry; D006801:Humans; D027522:Marrubium / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D019800:Phenol / Q000737:chemistry / Q000633:toxicity; D035261:Plant Components, Aerial / Q000737:chemistry / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D019800:Phenol; D000431:Ethanol; D020169:Caspases