Abstract
Cuphea aequipetala (C. aequipetala) has been used in Mexican traditional medicine since prehispanic times to treat tumors. In this paper, we evaluated the antiproliferative and apoptotic effect of the methanolic and aqueous extracts of C. aequipetala on several cancer cell lines including the B16F10 cell line of murine melanoma and carried a murine model assay. In vitro assay analyzed the effect in the cellular cycle and several indicators of apoptosis, such as the caspase-3 activity, DNA fragmentation, phosphatidylserine exposure (Annexin-V), and induction of cell membrane permeabilization (propidium iodide) in the B16F10 cells. In vivo, groups of C57BL/6 female mice were subcutaneously injected with 5x105 B16F10 cells and treated with 25 mg/mL of C. aequipetala extracts via oral. Aqueous and methanolic extracts showed a cytotoxic effect in MCF-7, HepG2, and B16F10 cell lines. The methanolic extract showed more antiproliferative effect with less concentration, and for this reason, the in vitro experiments were only continued with it. This extract was able to induce accumulation of cells on G1 phase of the cell cycle; moreover, it was able to induce DNA fragmentation and increase the activity of caspase-3 in B16F10 cells. On the other hand, in the murine model of melanoma, the aqueous extract showed a greater reduction of tumor size in comparison with the methanolic extract, showing an 80% reduction versus one of around 31%, both compared with the untreated control, indicating a better antitumor effect of C. aequipetala aqueous extract via oral administration. In conclusion, the in vitro data showed that both C. aequipetala extracts were able to induce cytotoxicity through the apoptosis pathway in B16F10 cells, and in vivo, the oral administration of aqueous extract reduces the melanoma tumoral mass, suggesting an important antitumoral effect and the perspective to search for effector molecules involved in it.
Highlights
Malignant melanoma is the most aggressive form of skin cancer [1, 2], because of its ability to metastasize to other organs such as lungs, liver, brain, or lymph nodes [3, 4]
In this work we evaluated the cytotoxic effect of C. aequipetala in several cancer cell lines, and carried an in vivo model to analyze the antitumor effects of the methanolic and aqueous extracts of this plant against murine melanoma B16F10 cells
The aqueous extract showed an important reduction of cell viability after 48h of exposure; the cytotoxic concentration that induces around 50% of cell death (CC50) in B16F10 was 0.364 mg/mL, in HepG2, was 0.212 mg/mL and in MCF-7 was 0.173 mg/mL (Figure 1(a))
Summary
Malignant melanoma is the most aggressive form of skin cancer [1, 2], because of its ability to metastasize to other organs such as lungs, liver, brain, or lymph nodes [3, 4]. Morbidity and mortality are primarily associated with metastatic disease and poor responses to most standard chemotherapies [8]. Due to these problems, efforts to find new and better alternatives to increase the survival of patients with melanoma are a necessity. Efforts to find new and better alternatives to increase the survival of patients with melanoma are a necessity From this perspective, plant-derived compounds have received considerable attention in recent years because of their pharmacological properties, such as cytotoxicity and chemotherapeutic activities in cancer [9, 10]. The BioMed Research International main drugs currently used for cancer treatment have been isolated from natural sources, which include taxol from Taxus species, vinblastine and vincristine from Catharanthus roseus, topotecan and irinotecan from Camptotheca acuminata, and etoposide and teniposide from Podophyllum peltatum [11]
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