Effect Of PRMT2 On The Proliferation Of Breast Cancer Cells

Objective: To investigate the effects of PRMT2 on the proliferation of human breast cancer MCF-7 cell and MDA-MB-231 cell, and to explore the potential mechanism of modulation of cyclin D1 by PRMT2.Methods: MDA-MB-231 cells were transicently transfected with pc DNA3.1/pc DNA3.1-PRMT2 vector to detect the CCND1 protein expression level with Western Blot. After establishing and identifying the tet-on expression system of PRMT2 packaged by the Lentivirus vector with 293 T cells, the MCF-7 and MDA-MB-231 cells were infected with the vectors to build stable cell lines. MTT and clone formation assay were applied to clarify the effects of over expression of PRMT2 on the proliferation of both stable cell lines. The expression of p-Akt、p-GSK-3β and CCND1 were detected by Western blot to investigate the potential mechanisms of PRMT2 modulating the proliferation of MCF-7 and MDA-MB-231 cells.Results:1. Western blot assay showed that CCND1 was decreased with transient over expression of PRMT2 in MDA-MB-231cells(p<0.05).2. Lentivirus expressive PRMT2 was successfully constructed and identified by sequencing. The titer(2.0 ×108 TU/ml) was assessed by Real Time PCR and the expression of PRMT2-3Flag was induced by Doxycycline hyclate and detected by Western blot in PRMT2 Lentivirus with 239 T cells.3. Western Blot results showed PRMT2-3Flag proteins were expressed in both cells after transfection(p<0.05).4. MTT showed that the proliferation rates of both cells were decreased(p<0.05).5. Colone formation assay showed the capacities of forming clone of both cells were suppressed(p<0.05).6. Western Blotting results indicated that PRMT2 down regulated the expression of CCND1, p-Akt and p-GSK-3β in both cells(p<0.05).Conclusions:1. PRMT2 could depress the proliferation of breast cancer cells.2. The potential mechanism of PRMT2 suppressing the proliferation of breast cancer cells could be the modulation of p-Akt, p-GSK-3β and CCND1 by PRMT2.