Cytotoxicity of Propolis Extracts obtained using Dichloromethane and Hexane Solvent on Human Salivary Gland Tumor Cell Line.

Abstract

This in vitro study aimed to investigate the effect of propolis extracts from two different solvents on human submandibular salivary gland (HSG) tumor cell line. Propolis was extracted by dichloromethane (DCM) and hexane (HEX). Crude extracts were prepared from 6.25 to 200 µg/mL in Dulbecco's modified eagle medium without serum. Flavonoid and total phenolic contents of crude extracts were measured using a modified colorimetric method. The cytotoxicity was evaluated by 3-[4, 5-dimethylthiazol-2-yl]-2,5 diphenyl-tetrazolium (MTT) assay and lactate dehydrogenase (LDH) release assay. The statistics were analyzed by independent sample t-test. Propolis extracts obtained using DCM and HEX exhibited comparable % yield (38.58 and 38.25) and physical characteristics and different amounts of flavonoid (0.439 ± 0.02 and 0.250 ± 0.01 mg catechin/g sample) and total phenolic compounds (3.759 ± 0.03 and 1.618 ± 0.03 mg gallic acid equivalents/g sample). The DCM group at 25, 50, 100, and 200 µg/mL as well as the HEX group at 50, 100, and 200 µg/mL significantly displayed a decrease in % cell viability and an increase in % cytotoxicity, compared with the untreated control group (P < 0.05). The DCM group showed the half-maximal inhibitory concentration (IC50) of MTT (42.93 ± 2.70) and LDH (34.94 ± 0.22). The HEX group showed the IC50 of MTT (61.30 ± 5.39) and LDH (42.32 ± 1.00). Propolis extracts obtained using both DCM and HEX are effective to inhibit HSG viability. Regarding to the cell morphological observation, MTT and LDH assays, propolis extracts obtained using DCM and HEX exhibited the cytotoxic effect on HSG tumor cell line. Based on our knowledge, this research demonstrates the first preliminary result suggesting propolis as a natural product of choice for salivary gland cancer prevention and therapy.