Effects of Recombinant Human Erythropoietin on MPC-11 Myeloma Cells Cultured In Vitro / 中国实验血液学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effect of recombinant human erythropoietin (rhEPO) on myeloma cell line MPC-11 in vitro and its anti-tumor mechamism so as to provide the theoretic evidence for treatment of multiple myeloma by using rhEPO.</p><p><b>METHODS</b>MPC-11 cells were cultured in RPMI 1640 medium, then the MPC-11 cells in logarithmic growth phase were seed in 96 well culture plates at a density of 1×10/ml cells with 100 µl per well. For all the experiments, the control and rhEPO groups were set up, and the rhEPO group was divided into different concentration groups 20, 40, 60, 80, 100, 200 and 400 rhEPO U/ml. The cultured MPC-11 cells in each groups were collected at day 1, 2, 3, 4, 5 and 6, and the viability of MPC-11 cells was assayed by using CCK-8 method, the MPC-11 cell apoptosis was detected by flow cytometry with Annexin V/PI double staining, the levels of IL-6, IgG and kappa light chain in cell supernatant were detected by ILISA.</p><p><b>RESULTS</b>The apoptosis rate in MPC-11 cells treated with 200 U/ml of rhEPO at day 21 was significantly higher than that in control group(P<0.05); the apoptosis rate in MPC-11 cells treated with 400 U/ml of rhEPO at day 10, 15, 21 was higher than that in control group at same time points, respectively (P<0.05), suggesting that the MPC-11 cell apoptosis rate was enhanced with increase of rhEPO concentration and prolonging of culture time(P<0.05), the IL-6 level in supernatant of cell treated with 400 U/ml of rhEOP for 15 days was lower than that in control group(P<0.05), while the IL-6 level in supernatant of cells treated with 100, 200, 400 U/ml of rhEPO for 21 days decreased (P<0.05); The levels of IgG and kappa light chain in supernatant of cells treated with 400 U/ml of rhEPO for 21 days all were significantly lower than those in control group(P<0.05).</p><p><b>CONCLUSION</b>The rhEPO can induce apoptosis of MPC-11 cells with concentration- and time-dependent manner, and can reduce levels of IgG, kappa light chain secreted by MPC-11 cells with time-dependent manner.</p>