Hernández John Fredy, Urueña Claudia Patricia, Cifuentes Maria Claudia, Sandoval Tito Alejandro, Pombo Luis Miguel, Castañeda Diana, Asea Alexzander, Fiorentino Susana
Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Pontificia Universidad Javeriana, Carrera 7 N. 43-82 Building 52, Office 608, Bogotá, Colombia.
Grupo de Farmacología Vegetal, Fundación Universitaria Juan N. Corpas, Carrera 111N. 159A-61, Bogotá, Colombia.
J Ethnopharmacol. 2014 May 14;153(3):641-9. doi: 10.1016/j.jep.2014.03.013. Epub 2014 Mar 15.
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).
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).
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.
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.
在加勒比地区、中美洲和南美洲，民间医学使用蒜藜芦（商陆科）的水提取物和醇提取物治疗白血病和乳腺癌。在此，我们使用转移性乳腺腺癌模型（4T1）验证了蒜藜芦提取物的生物活性。
通过评估4T1细胞的细胞增殖、细胞集落生长能力和细胞凋亡（半胱天冬酶-3活性、DNA片段化和线粒体膜电位）来确定蒜藜芦提取物的生物活性。蒜藜芦提取物使用IC₅₀浓度（29μg/mL）及以下的2种稀释液，阿霉素使用0.27μg/mL（阳性对照），二苄基二硫醚使用2.93μg/mL（IC50提取物标记化合物）。通过液相色谱-质谱联用仪分析蛋白质组学评估结果。通过逆转录-聚合酶链反应确认蛋白质水平表达。通过酶促测定法测量葡萄糖和乳酸水平。在BALB/c小鼠中确定半数致死量，并在移植了绿色荧光蛋白标记的4T1细胞的小鼠中评估抗肿瘤活性。通过腹腔注射用蒜藜芦提取物治疗小鼠（182mg/kg，相当于半数致死量的1/8及以下的2种稀释液）。
蒜藜芦提取物在体外可诱导4T1细胞凋亡、半胱天冬酶-3激活、DNA片段化且不伴有线粒体膜去极化，并降低细胞集落生长能力。此外，糖酵解酶表达的变化导致葡萄糖摄取和乳酸生成减少。该提取物还可促进移植了绿色荧光蛋白标记的4T1细胞的BALB/c小鼠的乳腺原发性肿瘤消退。
蒜藜芦叶和茎的提取物在小鼠乳腺癌模型中可诱导体外细胞死亡和体内肿瘤消退。我们的结果部分验证了蒜藜芦在乳腺癌治疗中的传统用途，揭示了一种设想蒜藜芦生物活性的新方法。该提取物对糖酵解途径酶的作用有助于解释其抗增殖和抗肿瘤活性。
