Effects of Erythropoietin and Erythropoietin Receptor on Human Brain Glioma Using siRNA Expression Vector

摘要

To investigate the effects of erythropoietin (EPO) and EPO receptor (EPOR) on the proliferation, apoptosis and migration of human brain glioma using the small interfering RNA (siRNA) expression vector.Human EPO siRNA plasmid expression vector was constructed with liposome Lipofectamine 2000, liposome Viafect and NEPA21 Electro-Kinetics Transfection system, respectively, to transfect human glioma U251 cells and the transfection rate was calculated with fluorescent microscope EPOR siRNA Lentivirus expression vector was used to transfect U251 cells.The proliferation, migration and apoptosis of U251 cells were measured with colony formation, scratch assay and flow cytometry, respectively.The optimal concentration of EPO for U251 treatment was 18.7 IU/L by MTT EPO increased the proliferation and migration and decreased the apoptosis of U251 cells.Among these systems that successfully constructed EPO siRNA expression vectors, the Electro-Kinetic Transfection system produced significantly higher transfection rate than liposome Lipofectamine 2000 and Viafect.The transfection rate within same transfection system increased following the increase of vector concentration and the Electro-Kinetic Transfection system increased the most, reaching the highest transfection rate of 86.9%.The successfully constructed EPOR siRNA expression vector LVEP0R produced the multiplicity of infection (MOI) of 100 for U251 cells.These results provide basis for early diagnosis and therapy of glioma.