ANTIPROLIFERATIVE EFFECTS OF PLANTS EXTRACTS FROM THE CERRADO BRAZILIAN BIOME IN PHEOCHROMOCYTOMA PC-12 CELL LINE

Program: Abstracts - Orals, Featured Poster Presentations, and Posters
Session: SUN 17-28-Adrenal Tumors & Pheochromocytoma
Basic/Clinical
Sunday, June 16, 2013: 1:45 PM-3:45 PM
Expo Halls ABC (Moscone Center)

Poster Board SUN-26
Beatriz Taynara Araujo dos Santos*1, Petra Ariadne Araujo1, Damaris Silveira1, Francisco de Assis Rocha Neves2, Perola Dias3 and Adriana Lofrano Alves Porto4
1Faculty of Health Sciences - University of Brasilia, Brasilia, Brazil, 2Univeristy of Brasilia, Brasilia DF, Brazil, 3FAculty of Health Sciences - University of Brasilia, Brasilia, Brazil, 4University of Brasilia, Brasilia DF, Brazil
Introduction: Pheochromocytomas are highly vascularized tumors arising from adrenal medulla. The majority of cases are benign, but malignant forms have been described. Surgical resection is the only definitive healing for benign tumours, while malignant tumors still have no effective treatment. Symptoms from malignant disease are usually managed with cytotoxic chemotherapy and adrenergic blockers, but several new therapeutic targets have been studied recently. Most chemotherapy commercialized drugs are obtained by semi-synthesis from chemical compounds of plants and fungi, and Brazilian biodiversity could support on new drugs development for this pathology. Methods: The PC-12 Adh was purchased from BCRJ (Banco de CÚlulas do Rio de Janeiro), and maintained in HAM-F12 Medium supplemented with horse serum, fetal bovine serum, sodium bicarbonate, 10% penicillin/streptomycin stored at 37°C and 5% CO2.  For the experiments, cells were seeded at a density of 5 x103 cell/well in 96-well plates, in triplicate. After 24h of incubation, cytotoxicity assay was performed after 24h and 48h of exposition to 500 and 1000μg/μL of each fraction (aqueous, ethanolic and hexanic) of 18 extracts obtained from Cerrado plants diluted in cell culture medium.  The extracts that demonstrated cytotoxic activity were submitted to a dose response curve assay with concentrations varying from 125 to 2000ug/uL. After treatment, cell viability was measured using MTT (3-(4,5-dimethylthiazole-2-yl)-2,5- diphenyltetrazolium bromide) colorimetric assay. Both experiments were performed on three independent analysis. Data were analyzed on GraphPad Prism 5® by one-way ANOVA with Tukey pos-hoc test for cytotoxicity assays, and non-linear regression and IC50 determination. Flow cytometry and genic expression experiments are in progress. Results: The species Allamanda blanchetti, Erytroxylum subrotundum, Genipapa Americana and Sapinus saponaria demonstrated a proliferation inhibitory activity ≥ 50% on cytotoxicity assays and showed an adequate reproducibility on dose response assays and had their IC50 calculated. The IC50 is used for flow cytometry and genic expression experiments to evaluate the cell death profile and possible effects on apoptose signaling pathways. Conclusion: On this biological trial of Cerrado Biome, 5 of 18 plant extracts demonstrated the ability to control pheochromocytoma cell line proliferation. Our findings suggest the presence of active chemical compounds with cytotoxic activity, corroborating bioprospection studies for the development of new antiproliferative drugs.

Nothing to Disclose: BTADS, PAA, DS, FDARN, PD, ALAP

*Please take note of The Endocrine Society's News Embargo Policy at http://www.endo-society.org/endo2013/media.cfm

Sources of Research Support: Conselho Nacional de Pesquisa - Brasil (CNPq - grant number 564658-2010-3)