Clinical Efficacy Of Thalidomide Combined With Chemotherapy In The Treatment Of Leukemia

Background and Purpose:Acute leukemia（AL）is a kind of malignant clonal tumor with high incidence and difficult clinical treatment.The current clinical treatment of AL is mainly chemotherapy,which can alleviate the condition of patients to a certain extent,but the curative effect is limited and it is easy to relapse,and some patients develop refractory AL.In order to improve the therapeutic effect of refractory AML patients and reduce the recurrence of the disease,it is very important to actively seek effective clinical treatment methods.Thalidomide（TLD）can inhibit the hyperplasia of blood vessels and improve the regulation of immune function,and has achieved good efficacy in clinical trials.This study was conducted to observe the combination of thalidomide and chemotherapy in the treatment of refractory AML,in order to provide a more effective treatment plan for refractory AML patients.Method:1.In this study,TLD combined with chemotherapy was selected as the treatment regimen,and the clinical effect of this regimen on refractory AML patients was analyzed and studied.Meanwhile,the effects of TLD combined with chemotherapy on the secretion of vascular endothelial growth factor（VEGF）and the expression of P-glycoprotein（P-g P）in refractory AML cells and HL-60/VCR cells were investigated.2.Cultivate HL-60/VCR cells and set up groups A,B,C,D,E,F,G and H.No drug was added to group A,and different doses of TLD（75ug/ml,150ug/ml,300ug/ml）were added to groups B,C,and D.E,F,G groups were added with different doses of As₂O₃（1umol/l,5umol/l,25umol/l）.Group H simultaneously added 300ug/ml TLD and 25umol/l As₂O₃.Bone marrow fluid was collected from patients,and BMMNC cells Group a did not add any drugs;group d added TLD 300ug/ml,group f added As₂O₃ 5umol/l,group h added300ug/ml TLD and 25umol/l As₂O₃ at the same time were isolated from it and cultured in vitro.Groups a,d,f,and h were set.3.Double antibody sandwich method was used to detect the effects of TLD and As2O3 at different time and drug concentration on VEGF concentration in HL-60/VCR cell and BMMNC cell supernatant according to Elisa kit.4.The effects of different time and different concentrations of TLD and As2O3 on the expression of P-g P on the surface of HL-60/VCR cells and BMMNC cells were detected by flow cytometry.5.Statistical methods:SPSS 11.5 software was used for the analysis of experimental data,analysis of variance was used for the comparison of multiple samples,and experimental data were expressed by mean standard deviation or percentage.The T-test and Chi-square test were performed in pairs respectively,and P<0.05 indicated significant difference.Result:1.In chemotherapy group,there were 2 cases of CR,4 cases of PR,14 cases of NR,8 cases of CR,6 cases of PR,and 6 cases of NR in the combination group.The effective rate of the combination group was 70%,which was higher than 30%of the control group（P<0.05）.2.The adverse reactions in the chemotherapy group were mainly nausea and vomiting,hair loss,liver and kidney function damage,etc.In the combination group,after increasing TLD,adverse reactions such as drowsiness and abdominal distension occurred.There was no significant difference in the occurrence of adverse reactions between the two groups of refractory AML patients（P>0.05）.3.The plasma VEGF concentrations of refractory AML patients and healthy subjects were（339.71±24.46）ng/L and（122.70±15.30）ng/L,respectively,and there was a significant difference between the two groups（t=33.638,P<0.01）.4.Compared with the concentration of VEGF in the supernatant of HL-60/VCR cells cultured for 48h,the concentration of VEGF in the supernatant of HL-60/VCR cells cultured for 72h in groups C,D,E,F,G and H was decreased（P<0.01）.Comparison of undosed group,single drug group and combined drug group:After 48h culture,compared with group A,VEGF concentration in the supernatant of HL-60/VCR cells in groups B,C,D,E and F had no significant difference（P>0.05）,while VEGF concentration in the supernatant of HL-60/VCR cells in group G was decreased（P<0.05）.The concentration of VEGF in the supernatant of HL-60/VCR cells in group H was decreased（P<0.01）.After72h of culture,VEGF concentration in the supernatant of HL-60/VCR cells in group B had no significant difference compared with group A（P>0.05）,while VEGF concentration in the supernatant of HL-60/VCR cells in groups C,D and E was decreased（P<0.05）.VEGF concentration in supernatant of HL-60/VCR cells in groups F,G and H was decreased（P<0.01）.Comparison between the combination group and the high-dose monotherapy group:After 48h culture,compared with group H,VEGF concentration in the supernatant of HL-60/VCR cells in group D was increased（P<0.01）,and VEGF concentration in the supernatant of HL-60/VCR cells in group G was not significantly different（P>0.05）.After72h culture,compared with group H,VEGF concentration in supernatant of HL-60/VCR cells in groups D and G was increased（P<0.05）.5.There was no significant difference in the expression rate of P-GP on the cell membrane of HL-60/VCR at 48h compared with that of large VP-g P at 72h among all groups（P>0.05）.After 48h and 72h,there was no significant difference in the expression rate of P-GP on the cell membrane of HL-60/VCR in the no-drug group,the monotherapy group and the combined treatment group（P>0.05）.6.Compared with the supernatant of BMMNC cells cultured for 48h,VEGF concentration in the supernatant of BMMNC cells cultured for 72h in groups d,f and H was decreased（P<0.01）.Compared with group a,the concentration of VEGF in the supernatant of BMMNC cells in groups d,f and h decreased after 48h and 72h culture（P<0.01）.Comparison between the combined treatment group and the single treatment group:after48h and 72h culture,VEGF concentration in the supernatant of BMMNC cells in groups d and f was increased compared with group h（P<0.01）.7.There was no significant difference in the expression rate of P-GP on BMMNC cell membrane between 48h and 72h among all groups（P>0.05）.After 48h and 72h culture,there was no significant difference in the expression rate of P-GP on the cell membrane surface of BMMNC in groups d,f and h compared with group a（P>0.05）.Conclusion:1.TLD combined with chemotherapy can improve the treatment efficiency of refractory AML patients without increasing the adverse reaction rate,which is safe and effective.2.The plasma VEGF concentration of refractory AML patients is higher than that of healthy people,suggesting that leukemia cells can secrete VEGF in large quantities.3.Both TLD and As2O3 can inhibit the ability of HL-60/VCR and BMMNC cells to secrete VEGF,and the combined use has a more obvious inhibitory effect.4.TLD and AS2O3 did not affect the expression of P-g P on HL-60/VCR and BMMNC cell membranes.5.In BMMNC cells isolated from refractory AML patients,the content of serum VEGF and the expression rate of P-g P on cell membrane surface can be used as indicators to monitor the disease and prognosis of refractory AML patients.