Anti-metastatic evidence of Tessaria absinthioides aqueous extract on a model of murine melanoma.

SOSA LOCHEDINO A; MONDACA J; HAPON MB; GAMARRA LUQUES C

Mendoza

Congreso; XXXIX Reunión Anual de la Sociedad de Biología de Cuyo.; 2021

Sociedad de Biología de Cuyo

Melanoma is one of the most aggressive and therapy-resistant cancers. Therefore, finding new treatments to improve patient outcomes is an ongoing effort. In our laboratory, we previously demonstrated that Tessaria absinthioides aqueous extract (TaAE) evidence in vitro cytotoxicity and exert in vivo antimelanoma effects delaying tumor detection, reducing tumor doubling time and diminishing the final volume of subcutaneous tumors. In the current work, we hypothesized that TaAE interfere with the melanoma metastasis and our goal is to quantify the effects on the in vitro migration and adhesion; as well as, to evidence the in vivo effects on metastasis. The B16-F10 (murine metastastatic melanoma) was the cell line used in this work; the cell migration after 24 hs was studied by the wound-healing assay; and to evaluate adhesion, attached cells into 6 well plates were quantified by trypan blue at 3, 6, 12 and 24 hs after they were plated. Then, the in vivo process of metastasis were evaluated by injection of 1x105 cells into the tail lateral vein of C57BL6/j mice; and pulmonary tumors were counted 21 days later. The in vitro studies were performed at doses of 10 µg/mL of TaAE, which is the higher dose that no affect proliferation after 24 hs (MTT determined). To treat the animals, TaAE was administrated in drinking water at 150 mg/kg/day. In the wound healing assay, we examined cell migration in response to the mechanical scratch wound in the absence or presence of TaAE. The scratch was photographed at 0 and 24 h later, measured by the image J® software, and then the measurements were compared by the Student´s T test. In the controls, the wound area were closed in a 13.75 ± 2.3 %. While in the TaAE treated wells, migration was significantly reduced (p˂0.05) and scratch was closed only in a 0.21 ± 2.7 %. On the other hand, when cells were treated with TaAE, the adhesion to the plate was also significantly reduced in all analyzed periods (Student´s T test, p˂0.05). In the control, no treated, group 49,22 ± 1,1 % of cells were attached at 3 hs; 85,08 ± 13,6 % at 6 hs; 118,3 ± 7,8 % at 12 hs; and 121,4 ± 9,3 % 24 hs later. In the TaAE treated wells, the attached cells were 1.65 ± 0.4%; 12.4 ± 0.2%; 69.37 ± 3.24% and 92.64 ± 3.1%, respectively. Finally, the in vivo study performed on C57BL6/j mice (n=10, per group) confirm the previous studies obtained in vitro; the TaAE in drinking water, at doses of 150 mg/kg/day, significantly reduce (Student´s T test, p˂0.05) the pulmonary metastasis from 161.6 ± 4.3/animal in the control group to 28 ± 1.8/animal in the treated group. Altogether, the presented results support the anti-metastatic activity of TaAE and open a new window in the study of the anticancer properties related to Tessaria´s extract, with the final objective of to promote the use of this botanical compound as complementary agent in the cancer treatment.