The Effect Of PRMT2 On Akt Phosphorylation Through Ppar? In Breast Cancer MCF-7 Cells

Objective: To explore the effect of PRMT2 on Akt phosphorylation through PPAR? and to preliminarily study the influence of PRMT2 on breast cancer stem cells in MCF-7 cells, in order to provide new insights into hormonal therapy of breast cancer.Methods: MCF-7-PRMT2 cells and MCF-7 cells were utilized in the present study to investigate the effect of PRMT2 on Akt phosphorylation through PPAR? and the influence of PRMT2 on breast cancer stem cells.Our experiments including four parts:(1) Western Blotting analysis was used to analyze the impact of PRMT2 overexpression on p-Akt?p-GSK-3??CyclinD1?PPAR? expression in MCF-7 cells at the protein level;(2) The interaction between PPAR? and PRMT2 in MCF-7 cells was determined by immune-coprecipitation;(3) Western Blotting analysis was used to detect the impact of PRMT2 overexpression and PPAR? inhibition on p-Akt protein levels;(4) Flow Cytometry was carried out to evaluate the effect of PRMT2 on breast cancer stem cells(CD44~+/CD24-).Results: 1. Western blot analysis showed that the protein levels of p-Akt?p-GSK-3??CyclinD1 in the MCF-7 cell line were decreased upon overexpression of PRMT2, while PPAR? protein levels had no obvious change.2. Co-immunprecipitation revealed that PRMT2 interacted with PPAR? in MCF-7 cells.3. Western blot analysis indicated that PPAR? inhibition increased the protein levels of p-Akt in the MCF-7 cell line, while had no changes in PRMT2 protein levels,on the basis of it, overexpression of PRMT2 could decrease p-Akt protein levels.In addtion, inhibition of PPAR? could increase p-Akt protein levels on the basis of overexpression of PRMT2.4. Flow cytometry results revealed that compared with the control group, the MCF-7 cell line with stable high level expression of PRMT2 had a relatively lower rate of breast cancer stem cells( CD44~+ /CD24~–), with statistical significance(P<0.05).Conclusion: 1. In the MCF-7 breast cancer cell line, PRMT2 could decrease the phosphorylation level of Akt Partially through the combination with PPAR?.2. Overexpression of PRMT2 could lower the CD44~+/CD24~– subpopulation ratio in MCF-7 cells.