Comparison of the effect of epigenetical treatment in 2D and 3D cholangiocarcinoma tumor models / 中华肝胆外科杂志

ABSTRACT

Objective To investigate the effects of epigenetic drugs on 2D- and 3D-cultured cholangiocarcinoma cells in vitro.Methods In this study,we have built compact and round TFK-1 spheroid in poly-HEMA coated 96-well plate and determined the effects of epigenetical drugs on 2D and 3D cultured cholangiocarcinoma cellsTFK-1.Viability of 2D and 3D cells model was examined by WST assay and FDA/PI staining. Using methylation-specific PCR analysis,we demonstrated the changes of methylation status of promoters regarding three tumor suppressor genes APC,E-Cadherin,and p16 INK4A.Results The average diameters of compact and round TFK-1 spheroids were in the range of 350-400 μm.The TFK-1 spheroid cells were more resistant to the epigenetic drugs and demonstrated a 11.2155-fold higher IC50 values for hydralazine and valproic acid than the same cells grown as monolayer. Higher doses of epigenetic drugs were needed to reverse the hypermethylation status in 3D cultured cells than 2D cells; however,the parallel dosage - dependent characteristic did not show in the 3D spheroid group.Conclusions Taken together,we established a 3D culture model of human cholangiocarcinoma epithelial spheroid.The 3D spheroid cells are more complex than the 2D monolayer cells and their unique characteristics are able to affect the consequences of epigenetic therapy.The 3D spheroid is a promising model for the research of epigenetic therapy.