Evaluation of the effect of epigallocatechin-3-gallate in cultured human HepG2 cell genomic instability in three dimensions (3D)

• Grant number: 14/12262-2
• Support Opportunities: Regular Research Grants
• Start date: November 01, 2014
• End date: December 31, 2016
• Field of knowledge: Biological Sciences - Genetics - Mutagenesis
• Principal Investigator: Lusânia Maria Greggi Antunes
• Grantee: Lusânia Maria Greggi Antunes
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

The use of the 3Rs (Reduction, Refinement, and Replacement) was proposed as an alternative for replacing the use of animals in research in order to minimize the number of animals used and to refine the experimental design so as to cause the least possible amount of pain. The importance of applying new alternative methods to the use of animals has led to the creation of new in vitro methods. It has been suggested that a three-dimensional (3D) cell culture, in contrast to the conventional bidimensional (2D) culture, more accurately represents the original microenvironment of cells contained in tissues, thereby more accurately reflecting the in vivo response. One of the most widely used in vitro systems is multicellular spheroids, which has been sufficiently well characterized as reliably reproducing the microenvironment of a tumor. In recent years, many studies have suggested that natural products and dietary bioactive compounds with potential chemopreventive effects can induce epigenetic changes. Many studies have demonstrated the antitumor activity of epigallocatechin-3-gallate through epigenetic changes in a conventional 2D culture. However, there are no studies demonstrating the epigenetic effects of epigallocatechin-3-gallate in a 3D cell culture. Thus, our objective is to evaluate the effect that epigallocatechin-3-gallate activity has on the cellular and morphological integrity of multicellular tumor spheroids, three-dimensional cell migration and invasion, and epigenetic changes in the genes and proteins involved in cell proliferation in HepG2 cultivated as multicellular tumor spheroids in a three-dimensional culture (3D). (AU)