Petiveria alliacea extracts uses multiple mechanisms to inhibit growth of human and mouse tumoral cells

Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Universidad Javeriana, Bogotá, Colombia.
BMC Complementary and Alternative Medicine (Impact Factor: 2.02). 12/2008; 8(1):60. DOI: 10.1186/1472-6882-8-60
Source: PubMed


There is ethnopharmacological evidence that Petiveria alliacea can have antitumor activity; however, the mechanism of its cytotoxic activity is not well understood. We assessed multiple in vitro biological activities of an ethyl acetate soluble plant fraction over several tumor cell lines.
Tumor cell lines were evaluated using the following tests: trypan blue exclusion test, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide], flow cytometry, cytoskeleton organization analysis, cell cycle, mitochondria membrane depolarization, clonogenicity test, DNA fragmentation test and differential protein expression by HPLC-Chip/MS analysis. F4 fraction characterization was made by HPLC-MS.
Petiveria alliacea fraction characterized by de-replication was found to alter actin cytoskeleton organization, induce G2 cell cycle arrest and cause apoptotic cell death in a mitochondria independent way. In addition, we found down regulation of cytoskeleton, chaperone, signal transduction proteins, and proteins involved in metabolic pathways. Finally up regulation of proteins involved in translation and intracellular degradation was also observed.
The results of this study indicate that Petiveria alliacea exerts multiple biological activities in vitro consistent with cytotoxicity. Further studies in animal models are needed but Petiveria alliacea appears to be a good candidate to be used as an antitumor agent.

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    • "Cell colony growth capacity assays were performed as previously described (Urueña et al., 2008). "
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    ABSTRACT: Folk medicine uses aqueous and alcoholic extracts from Petiveria alliacea (Phytolaccaceae) in leukemia and breast cancer treatment in the Caribbean, Central and South America. To evaluate in vitro and in vivo biological activity of a P. alliacea ethyl acetate fraction derived from leaves and stems in a metastatic breast adenocarcinoma model (4T1). P. 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. Doxorubicin (0.27µg/mL) was used as positive control and P. alliacea concentrations at the IC50 (29µg/mL) and 2 dilutions below. Dibenzyl disulfide (2.93µg/mL IC50) used in cytotoxic assays as 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 P. alliacea fraction via I.P (182mg/Kg corresponding to 1/8 of LD50 and 2 dilutions below). P. 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. A fraction from P. 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 P. alliacea in breast cancer treatment, revealing a new way of envisioning P. alliacea biological activity. The fraction effect on the glycolytic pathway enzymes contributes to explain the antiproliferative and antitumor activities.
    Journal of ethnopharmacology 03/2014; 153(3). DOI:10.1016/j.jep.2014.03.013 · 3.00 Impact Factor
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    • "P2Et fraction and doxorubicin cytotoxic effects on normal and tumor cells were evaluated using trypan blue and methylthiazol tetrazolium (MTT) assays (Sigma-Aldrich, Saint Louis, MO) as previously reported [17]. Cell viability was assessed with a trypan blue dye exclusion test. "
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    BMC Complementary and Alternative Medicine 04/2013; 13(1):74. DOI:10.1186/1472-6882-13-74 · 2.02 Impact Factor
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    • "The cytotoxic effects of the fractions and conventional chemotherapeutic drugs (doxorubicin, etoposide, vincristine and taxol) were evaluated using normal and tumor cells by means of trypan blue and the methylthiazol tetrazolium (MTT) assay, as previously reported [14]. The P2Et fraction was dissolve in ethanol and the corresponding vehicle was used as a negative control. "
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