The Role Of EPO Promoted The Proliferation Of Glioblastoma Cells Via Akt Pathway In Vitro

ObjectiveThis study will explore whether erythropoietin(EPO) plays a vital role in promoting proliferation through Akt and CyclinD1 in glioblastoma.MethodThe U87 cells were cultured in vitro and divided into three groups: control group treated with equal value PBS; EPO treatment group cultured with a dose of 0.1U/ml EPO for seven days; EPO+ Akt inhibitor group treated with EPO(0.1U/ml) and Akt inhibitor(81.3μM) for seven days. The proliferation rate was measured by Cell Counting Kit-8(CCK-8) assay, Double time and Clone formation,CyclinD1 expression was detected by Reverse Transcription-Polymerase Chain Reaction( RT-PCR), Western blotting(WB) and Cell immunofluorescence(IF), respectively. The cell cycle was analyzed by flow cytometry.ResultsCompared with those of control group, the proliferation of U87 cells was obviously increased by EPO( P CCK8 3/5/7day=0.020/0.028/0.007,P Double time =0.0269,P flow cytometry=0.0028, P Clone formation= 0.0010), and the mRNA and protein levels of CyclinD1 were significantly enhanced(PRT-PCR=0.0186,P P-Akt =0.0020,Pβ-Cantenin =0.0026,P CyclinD1 =0.0022,PIF=0.0099). Compared with those of EPO treatment group, Akt inhibitor inhibited the mRNA and protein levels of CyclinD1 induced by EPO(PRT-PCR=0.0029,P P-Akt <0.0001,P β-Cantenin =0.0003,P CyclinD1 <0.0001,PIF=0.0007), U87 cells were blockaded in G1 stage(P=0.0036), and the proliferation of ability was significantly reduced(PCCK83/5/7day=0.000/0.001/0.000,P Double time =0.0010, P Clone formation=0.0017). Conclusion Erythropoietin may significantly promote the proliferation of glioma U87 cells through upregulated the expression of cyclin D1.