Citotoxic effects of ellagic acid on 2D and 3D cell culture

SGARIGLIA, M.A.; SOBERÓN, J.R.; ANDINA, M.L.; VATTUONE, M.A.

Amsterdam

Congreso; 6th International Conference and Exhibition on Traditional & Alternative Medicine; 2016

OMICS International Conference

The plant-derived compounds have attracted considerable attention as alternative cancer remedies for enhancing prevention and treatment because of their low toxicities and low side effects. Ellagic acid (EA) is a plant phenolic constituent that elicits anticarcinogenic effects by inhibiting tumor cell proliferation, inducing apoptosis, and disturbing inflammation, as was evidenced in multiple studies. However, its in vitro activity in three-dimensional culture models has been little studied. The tumor microenvironment in a 3D cell culture presents a heterogeneity like the in vivo non vascularized microtumor, in terms of gradients of oxygen, nutrients, and metabolites. Cell spheroids from F88.2 fibroblastic human cell line were obtained by Liquid Overlay method: 20,000 cells were cultivated in 96 well-plate previously coated with 1 % agarose, and incubated for 72 h (DMEM/5%BFS). 0.1, 1.0 and 2.5 µM EA treatments were included in DMEM in 24 well-plate with ten spheroids/well, during 3 h, 37 ºC, 5%CO2; 0.05% DMSO was the control. Trypan Blue exclusion test and MTT assay were used after 24 h total incubation to determine cell viability. EA was tested in 2D culture for comparative purposes. Fibroblasts on monolayer culture showed altered morphology, showing apoptotic signs. MTT assay showed that F88.2 cells were sensible to EA by reducing the 75% viability at 1 µM.Spheroids showed no significant susceptibility to EA considering the membrane integrity and metabolic activity. This results suggested that antiproliferative activity of AE decreased in a complex system as F88.2 cell spheroids. Others experiments with different cell lines will be made.