Investigation Of The Function Of EZH2 Interacting Protein In Breast Cancer Cells

Objective: In this research,we aimed to verify the interaction between EZH2 and Mnc2,and explore the functions and mechanisms of EZH2 and Mnc2 in breast cancer cells.Methods : Co-immunoprecipitation and Western Blot method were employed to verify EZH2 interaction protein Mnc2.Mnc2-overexpression and-silencing breast cancer cell lines were obtained by transfection or lentivirus mediated infection methods.The regulation of H3K27me3 level by Mnc2 was tested through western Blots methods using anti-H3K27me3 antibody as first antibody.The cell cycle regulation function of Mnc2 was determined by cell number count methods in the Mnc2 gain of function and loss of function cell models.Results: The interaction of EZH2 and Mnc2 was verified by Co-IP.We constructed MDA-MB231-teton-HAMnc2 cell line that high level of HAMnc2 expression was detected by anti-HA antibody.In this cell line,we found that overexpression of Mnc2 could promote cell proliferation and that effect could be inhibited by EZH2 inhibitor De Zep.Furthermore,T47D-shMnc2 cell line was constructed to verify the cell cycle regulation function of Mnc2,and the results were in concord with Mnc2 overexpression cell models.In T47D-Tenton-shMnc2 cells,Mnc2 was silenced at 4 days after doxcycline induction,and the level of H3K27me3 were increased after Mnc2 silencing.Conclusion: Mnc2 was an EZH2 interaction partner,and Mnc2 could regulate EZH2-methylated H3K27me3 level.In breast cancer cells,we determined Mnc2 could promote cell proliferation,and it may be a potential cancer therapy target.