title: Acetonic extract of Buxus sempervirens induces cell cycle arrest, apoptosis and autophagy in breast cancer cells.
abstract: Plants are an invaluable source of potential new anti-cancer drugs. Here, we investigated the cytotoxic activity of the acetonic extract of Buxus sempervirens on five breast cancer cell lines, MCF7, MCF10CA1a and T47D, three aggressive triple positive breast cancer cell lines, and BT-20 and MDA-MB-435, which are triple negative breast cancer cell lines. As a control, MCF10A, a spontaneously immortalized but non-tumoral cell line has been used. The acetonic extract of Buxus sempervirens showed cytotoxic activity towards all the five studied breast cancer cell lines with an IC(50) ranging from 7.74 µg/ml to 12.5 µg/ml. Most importantly, the plant extract was less toxic towards MCF10A with an IC(50) of 19.24 µg/ml. Fluorescence-activated cell sorting (FACS) analysis showed that the plant extract induced cell death and cell cycle arrest in G0/G1 phase in MCF7, T47D, MCF10CA1a and BT-20 cell lines, concomitant to cyclin D1 downregulation. Application of MCF7 and MCF10CA1a respective IC(50) did not show such effects on the control cell line MCF10A. Propidium iodide/Annexin V double staining revealed a pre-apoptotic cell population with extract-treated MCF10CA1a, T47D and BT-20 cells. Transmission electron microscopy analyses indicated the occurrence of autophagy in MCF7 and MCF10CA1a cell lines. Immunofluorescence and Western blot assays confirmed the processing of microtubule-associated protein LC3 in the treated cancer cells. Moreover, we have demonstrated the upregulation of Beclin-1 in these cell lines and downregulation of Survivin and p21. Also, Caspase-3 detection in treated BT-20 and T47D confirmed the occurrence of apoptosis in these cells. Our findings indicate that Buxus sempervirens extract exhibit promising anti-cancer activity by triggering both autophagic cell death and apoptosis, suggesting that this plant may contain potential anti-cancer agents for single or combinatory cancer therapy against breast cancer.
authors.affiliation: Laboratoire de Biochimie Appliquée, Faculté des Sciences de la Nature et de la vie, Université de Béjaia, Béjaia, Algeria. ouardia.ait-mohamed@univ-paris-diderot.fr
authors.forename: Ouardia
authors.initials: O
authors.lastname: Ait-Mohamed
authors.forename: Valentine
authors.initials: V
authors.lastname: Battisti
authors.forename: Véronique
authors.initials: V
authors.lastname: Joliot
authors.forename: Lauriane
authors.initials: L
authors.lastname: Fritsch
authors.forename: Julien
authors.initials: J
authors.lastname: Pontis
authors.forename: Souhila
authors.initials: S
authors.lastname: Medjkane
authors.forename: Catherine
authors.initials: C
authors.lastname: Redeuilh
authors.forename: Aazdine
authors.initials: A
authors.lastname: Lamouri
authors.forename: Christine
authors.initials: C
authors.lastname: Fahy
authors.forename: Mohamed
authors.initials: M
authors.lastname: Rholam
authors.forename: Djebbar
authors.initials: D
authors.lastname: Atmani
authors.forename: Slimane
authors.initials: S
authors.lastname: Ait-Si-Ali
mesh_terms: D000096:Acetone / Q000737:chemistry*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects*; D000071186:Beclin-1; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000378:metabolism*; D031235:Buxus / Q000737:chemistry*; D002453:Cell Cycle / Q000187:drug effects*; D045744:Cell Line, Tumor; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry* / Q000494:pharmacology*; D000077022:Survivin
chemical_list: D051017:Apoptosis Regulatory Proteins; C491997:BECN1 protein, human; C107489:BIRC5 protein, human; D000071186:Beclin-1; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D051034:Inhibitor of Apoptosis Proteins; D008565:Membrane Proteins; D010936:Plant Extracts; D000077022:Survivin; D000096:Acetone