| Objective To explore the effects of recombinant human erythropoietin(rhEPO)on leukemic cell proliferation.Offer some experimental basis for the treatment of secondary anemia of leukemia with rhEPO for clinical application.Methods:Samples were collected from non-remission and recurrent acute leukemia patients(leukemia cells＞50%with the exception of M6)in Shanxi province hospital and the second clinical medical college of Shanxi medical university from October 26th,2006 to January30 th,2007.Thirty acute leukemia samples were investigated:15 men and 15 women, age from 5 to 63,the average age is 29±17 years old,including 21 acute myelogenous leukemia and 9 acute lymphocytic leukemia.①Take bone marrow or peripheral blood 2ml with EDTA anticoagulation pipe and wave evenly alone with the tubal wall,separate the mononuclearcell by lympholeukocyte separating medium,then abstract the mononuclearcell,clean cells two times with D-hank's Liq;Removal erythrocyte among mononuclearcell by erythrocyte lysate.Ft. 1X106/L dimensional MNC suspension with RPMI-1640 culture fluid and morphological observation and counting.②Subculturing mononuclearcell in culture table with 24 holes,according to preliminary experimental result and design,put rhEPO in each hole 10u,20u respectively,the holes with no rhEPO as blank,culture it in 37℃,5%CO2 incubaton.Collect cells at 24h,72h respectively.③Take cultural cell suspension 500μl from each hole,centrifuge it 1000r/min for 5 minutes,pour supernatant and misce bene cells.Take 20μl film preparation. morphological observation and counting.In vitro culture of leukemia cell was performed with additions of various concentrations of rhEPO.Treatments were terminated at 24h and 72h, respectively.Then each group of cells was incubated with FITC-1gG antibody to CD71.And TfR expression and DNA synthesis status were analyzed by flowcytometry.Results:Bone marrow cells of 30 acute leukemia patients have no marked change in morphology and counted number when incubated for 24h or 72h with different concentrations of rhEPO.P＞0.05.And have no significant deviation in contrast with each group.The expression of TfR by leukemic cells haven't increased significantly when incubated for 24h or 72h with different concentrations of rhEPO.P＞0.05,the same as S phase cells,also there was no marked change between AML and ALL.Conclusion:Acute leucemia patients express TfR all.Morphology and counted number and the expression of TfR by leukemic cells and S phase cells in 30 patients haven't increased significantly when incubated for 24h or 72h with different concentrations of rhEPO.In vitro leukemic cell have no significant proliferation with rhEPO.
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