A Petiveria alliacea standardized fraction induces breast adenocarcinoma cell death by modulating glycolytic metabolism.

ETHNOPHARMACOLOGICAL RELEVANCE: 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). MATERIALS AND 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. CONCLUSION: 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.

A fraction from Petiveria alliacea induces apoptosis via a mitochondria-dependent pathway and regulates HSP70 expression / Una fracción de Petiveria alliacea induce apoptosis dependiente de la mitocondria y regula la expresión de la proteína HSP70 / Uma fração de Petiveria alliacea induz apoptose dependente da mitocôndria e regula a expressão da proteína HSP70

Objective. To evaluate the biological activity of Petiveria alliacea extracts on tumoral cells in vitro. Materials and methods. P.alliacea fractions prepared by a bioguided purification protocol were characterized by their biological activities on two human tumoral cell lines. Morphological changes, cell viability, mitochondrial membrane depolarization, nuclear staining and activity on HSP70 were analyzed. Results. The present study demonstrates that P.alliacea fractions can induce apoptosis in a mitochondria-dependent pathway and down regulate HSP70 expression in vitro. The possible compounds present in P.alliacea responsible for the described biological activities were identified by dereplication methods. Conclusion. The present study provides new knowledge on the antitumoral properties of the plant species P. alliacea described in several ethnobotanical studies.

Objetivo. Evaluar la actividad biológica in vitro de extractos de Petiveria alliacea utilizando líneas celulares tumorales. Materiales y métodos. Las fracciones de P.alliacea fueron preparadas mediante un protocolo de purificación biodirigido. La actividad biológica fue caracterizada utilizando dos líneas celulares tumorales de origen humano, donde se analizaron los cambios morfológicos, la viabilidad celular, la actividad sobre la membrana mitocondrial y la fragmentación nuclear. Asimismo se evaluó la actividad de las fracciones sobre la expresión de la proteína HSP70. Resultados. El presente estudio muestra que las fracciones de P.alliacea inducen muerte celular por apoptosis dependiente de la mitocondria y además regulan negativamente la expresión de la proteína HSP70. Los compuestos que posiblemente son responsables de la actividad biológica presente en las fracciones de P.alliacea fueron identificados por dereplicación. Conclusión. El presente estudio contribuye a explicar en parte las propiedades antitumorales de la planta P.alliacea descritas en diversos escritos etnobotánicos.

Objetivo. Avaliar a atividade biológica in vitro de extratos de Petiveria alliacea utilizando linhagens de células tumorais. Materiais e métodos. Frações de P.alliacea foram preparadas utilizando um protocolo de purificação bio-dirigido. A atividade biológica foi caracterizada utilizando duas linhagens de células tumorais de origem humana, onde se analisaram as alterações morfológicas, a viabilidade celular, a atividade na membrana mitocondrial e a fragmentação nuclear. Também se avaliaram a atividade das frações sobre a expressão da proteína HSP70. Resultados. Este estudo mostra que as frações de P.alliacea induzem a morte celular por apoptose dependente da mitocôndria e também regulam negativamente a expressão da proteína HSP70. Os compostos que provavelmente são responsáveis pela atividade biológica presente nas frações de P.alliacea foram identificados por dereplicação. Conclusão. Este estudo ajuda a explicar, em parte, as propriedades antitumorais da planta P.alliacea descrita em vários escritos etnobotônicos.