| Objective To establish a 3D culture system of acute myeloid leukemia(non-APL)cells based on r BM system in vitro,and to explore the relationship between migration behavior and drug resistance of acute myeloid leukemia(non-APL)cells cultured in3 D,so as to provide a new idea for the treatment of acute myeloid leukemia(non-APL).Methods From June 2019 to December 2020,30 patients with newly diagnosed acute myeloid leukemia(non-APL)and 27 patients with relapsed drug-resistant acute myeloid leukemia(non-APL)were collected.The patients were divided into two groups: recurrent drug-resistant acute myeloid leukemia(non-APL)patients as the experimental group,newly diagnosed acute myeloid leukemia(non-APL)patients as the control group.The bone marrow fluid was obtained by using aseptic heparin anticoagulant tube,and the mononuclear cells in the bone marrow fluid were isolated and cultured by 3D technique.The 3D cultured cells of the two groups were treated with cytarabine(2 μ M).The cell migration trajectories were analyzed and drawn by Image J software,and quantified.The average cell migration rates of the experimental group and the control group before,48 hours and 96 hours after administration were compared between groups and within groups.Results(1)Cells in the experimental group and control group without cytarabine were continuously photographed under the microscope,and the software Image J was used to draw the cell migration trajectory and calculate the average migration rate of each group.The average cell migration rate of the experimental group was(57.28±17.63)μm/h,and that of the control group was(45.35±14.54)μm/h.The average cell migration rate of the experimental group was significantly higher than that of the control group(P < 0.01).(2)The leukemic cells treated with cytarabine for48 hours in the experimental group and the control group were photographed continuously under microscope and analyzed by Image J.It was found that the average migration rate of leukemic cells in the experimental group and the control group was(54.55 ±19.72)μm/h and(43.24±16.82)μm/h respectively,and the average cell migration rate in the experimental group was significantly higher than that in the control group(P < 0.05).(3)The results showed that the average cell migration rate was(53.53±18.82)μm/h in the experimental group and(42.53±15.84)μm/h in the control group 96 hours after addition of cytarabine.The average cell migration rate was significantly higher in the experimental group than in the control group 96 hours after addition of cytarabine(P<0.05).(4)The average cell migration rate of the experimental group and the control group was analyzed before and 48 and 96 hours after dosing.It was found that the average cell migration rate of the experimental group before dosing was(57.25±17.63)μm/h,48 hours after dosing was(54.55±19.72)μm/h,and 96 hours after dosing was(53.53±18.82)μm/h showed significant differences(P>0.05).There was no significant difference in the mean migration rate of cells in the control group at(45.35±14.54)μm/h before,(43.24±16.82)μm/h 48 hours after and(42.53±15.84)μm/h 96 hours after addition(P>0.05).Conclusion By establishing the in vitro 3D culture system of acute myeloid leukemia cells(non-APL)based on r BM system,it is found that the in vitro 3D culture system based on r BM system can well reproduce the bone marrow microenvironment and help us to study the relationship between migration behavior and drug resistance of acute myeloid leukemia(non-APL)cells.Using this culture system,we found that the cell migration rate of acute myeloid leukemia(non-APL)cells resistant to cytarabine increased significantly in 3D culture system in vitro,which provided new basic data for the future study of drug resistance of acute myeloid leukemia(non-APL),and helped to find new treatment options for acute myeloid leukemia(non-APL)and prolong the survival time of patients. |
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