Glycerol (GLY) inhibits hepatocellular carcinoma cell proliferation by reactive oxygen species (ROS) generation

CAPIGLIONI ALEJO; LORENZETTI FLORENCIA; QUIROGA, ARIEL DARÍO; PARODY J P; CARRILLO M C; MARINELLI R; CEBALLOS M P; ALVAREZ M L

Mar del Plata

Congreso; LXI Reunión Científica Anual de la Sociedad Argentina de Investigación Clínica (SAIC); 2016

SAIC

In previous studies our group reported antiproliferative and proapoptotic effects of GLY administration in an in vivo preneoplasic model. We postulate GLY as a possible ROS inductor that works as a signaling molecule after its metabolization to GLY phosphate (GlyP). Aim: To begin to study the antiproliferative mechanisms of GLY using hepatocarcinoma HepG2 cells. Methods and Results: MTT assay was used to determine the effects of GLY on HepG2cell line proliferation. The dose-response curve revealed that cell proliferation was diminished in a dose dependent manner (IC50=0.5% v/v). The primary cellular metabolite of GLY is GlyP. To analyze this metabolic fate of GLY, HepG2 cells were incubated at 37 0C for 72 h with 0.5% and 1% GLY and GlyP generation was measured by a colorimetric reaction. GlyP levels showed a47.5%* and 21.0%* increase in cells treated with GLY 0.5% and 1%, respectively, compared to control cells. It has been reported that subsequent GlyP metabolism induces ROS generation. To determine the possible occurrence of this phenomenon, after 72h of treatment, cells were incubated 30 min with 2′,7' Dichlorofluorescein diacetate (DCF-DA). Inside the cells, DCF-DA moleculeswere hydrolyzed by esterases, and reacted with intracellular ROS,producing a fluorescent product that can be measured fluorometrically.GLY caused a raise of 20%* in ROS production at both concentrations tested (*p