Influence of different culture methods in DNA methylation status of breast cancer MDA-MB-23 1 cells / 吉林大学学报(医学版)

ABSTRACT

Objective:To explore the influence of different cell culture methods in the genome-wide DNA methylation status of breast cancer MDA-MB-231 cells,and to clarify the relationship between genome-wide DNA methylation status and cell growth environment and the role of genome-wide DNA methylation status in the occurrence and development of tumor.Methods:The MDA-MB-231 cells were cultured with 2D and 3D cell culture models and mouse orthotopic transplantation model (Ti model)and collected, then DNA was extracted by DNA extraction kit and the genome-wide DNA methylation status of MDA-MB-231 cells after cultured with three different culture methods was detected by DNA methylation chip,then the value of beta,DiffScore and Delta_Beta of the CpG loci of each gene were calculated by applying Genomestudio software, and the differential methylation genes were screened by Genomestudio software and GO and Pathway analysis of these genes were performed in DAVID on-line analysis tool.Results:All 480 genes of the MDA-MB-231 cells showed significant differences in the degree of methylation in 3D and 2D models (P<0.05);86 448 genes in 3D and Ti models (P<0.05);90 005 genes in Ti and 2D models (P<0.05).The differential methylation genes in 3D and 2D,3D and Ti,and Ti and 2D models were enriched on the multicellular organismal development term and cell differentiation term (P<0.05);also on MAPK signaling pathway,cell adhesion molecules (CAMs),and regulation of actin cytoskeleton (P<0.05). Conclusion:There are differences in genome-wide DNA methylation status of MDA-MB-231 cells cultured in 2D, 3D cell culture and Ti models.