Screening And Identification Of EZH2 Interacting Proteins

Objective:To Screen and identify EZH2 interaction proteins in nasopharyngeal carcinoma(NPC)cells, for the further understanding of the EZH2 function during NPC formation and development.Methods:In this study, co-immunoprecipitation combined with MALDI-TOF or western blot were employed to explore the potential EZH2 interaction proteins. To obtained low nonspecific binding, the anti-flag antibody was used for CO-IP EZH2 interaction proteins from Flag-EZH2 overexpressed 5-8F cells. The lentivirus encoding flag fused EZH2 were packaged and used to infect 5-8F cells to obtain stable EZH2 overexpression cell line.Results:1.Produce a cell line that overexpressing EZH2, based on 5-8F cells. 2. Successfully identified 17 proteins with high fidelity to be potential EZH2 interaction proteins, via MALDI-TOF methods. 3. TF1 was further confirmed to be EZH2 interaction protein by CO-IP combined with WB.Conclusions:1.CO-IP combined with MALDI-TOF-MS wereemployed toidentifieda list of EZH2 interaction proteins.These results provide new hints for thefurtherunderstanding of EZH2 functionand the molecular mechanisms of nasopharyngeal carcinoma.2.TF1 protein interaction with EZH2 protein in nasopharyngeal carcinoma cells, and may beinvolved in the occurrence and development of NPC.