Estrogen receptor ERα36 gene silencing impacts the expression of growth-associated protein in PC12 cells

Abstract

Objective To investigate the effects of ERα36(a novel subtype of estrogen receptor alpha) on growth and proliferation in PC12 cells via examining the expression of growth-associated protein in differentiated PC12 cells after ERα36 gene silencing. Methods Transfection of ERα36-shRNA plasmid into PC12 cells was performed to establish the ERα36 gene silencing cells model(PC12-36L1, PC12-36L2). Immunocytofluorescence was used to examine the expression of ERα36, and Western blot was used to analyze the expression of PCNA, cyclinD1 and MAPK in the PC12 cells. Results ① ERα36 was expressed in both cell types(PC12-36C1, PC12-36L1 and PC12-36L2). Compared with PC12-36C1, PC12-36L2 cells(OD value were respectively 0.95±0.05, 0.78±0.10), PC12-36L1 cells significantly decreased expression of ERα36(OD value 0.47±0.12, P<0.01). ② Compared with PC12 and PC12-36C1 cells, PC12-36L1 cells were significantly higher expression of PCNA, CyclinD1 and p-MAPK(P<0.01)(OD value of PCNA, CyclinD1 and p-MAPK: PC12 cells were respectively 1.00±0.05, 1.00±0.11, 1.00±0.05, PC12-36C1 cells were respectively 1.09±0.15, 0.92±0.23, 1.12±0.08, PC12-36L1 cells were respectively 1.74±0.12, 2.20±0.25, 1.77±0.06). Conclusion ERα36 gene silencing can promote the growth and proliferation in PC12 cells.It suggests that the lower expression of ERα36 may be related to the diseases in nervous system such as brain tumor. Key words: Estrogen receptor gene, ERα36; Nerve cells; Growth-associated protein