【作者】 郑红艳；

【导师】 李慧慧；

【作者基本信息】 南昌大学 ， 内科， 2008， 硕士

【摘要】 目的:观察沙利度胺（TLD）和三氧化二砷（As₂O₃ ）对急性髓系白血病（AML）细胞分泌血管内皮生长因子（VEGF）和表达P-糖蛋白（P-gP）的影响。方法:收集AML患者和健康体检者的血液,肝素抗凝并离心后,吸取上层血浆置-20℃冰箱内冷冻保存。收集AML患者的骨髓液,用Ficoll淋巴细胞分离液分离出骨髓单个核细胞（BMMNC）。37℃、5%CO2、饱和湿度条件下培养细胞于RPMI l64O培养液中。HL-60/VCR细胞株的培养基中加入VCR以维持耐药性。TLD和As₂O₃以不同浓度加入培养体系中,同时设空白对照。用双抗体夹心酶联免疫法（ELISA）法检测血浆和细胞培养上清液中VEGF的浓度,用流式细胞术（FCM）检测细胞膜上MDR1/p-gp的表达率。结果:（1）AML患者血浆中VEGF浓度明显高于正常对照组（P<0.01）。（2）加入TLD和As₂O₃后,AML细胞和HL-60/VCR细胞株培养上清中VEGF浓度明显降低。（3）加入TLD和As₂O₃前后,AML细胞和HL-60/VCR细胞株的细胞膜上P-gP的表达率未见明显改变。（4）加入TLD和As₂O₃前后,AML细胞培养上清中VEGF的升高和细胞膜表面PgP的表达具有一定的正相关性。结论:（1）白血病细胞可以通过自分泌途径分泌VEGF。（2）TLD和AS2O3均可抑制HL-60/VCR和AML细胞分泌VEGF。（3）TLD和AS2O3均未能影响HL-60/VCR和AML细胞膜上P-gP的表达。（4）AML患者BMMNC培养上清中VEGF的升高和细胞膜表面PgP的表达具有一定的正相关性,二者联合检测可作为监测病情和判断预后的指标。更多还原

【Abstract】 Objective: To investigate the effect of thalidomide and As203 on the secretion of vascular endothelial growth factor（VEGF） and Pgp expression of the cell of AML.Methods:The patients with AML and healthy people in contrast group were analyzed. Heparin- anticoagulated blood were centrifuged to obtain plasma then conserved in -20℃fridge. The patients with AML were analyzed. Heparinized marrow was taken, and putted into Ficoll-isopaque solution to purified BMMNC.BMMNC and HL-60/VCR were both cultured in the RPMI 1640 medium, placed in 5% CO2 saturation and 37℃to proliferation.Added VCR to the medium of HL-60/VCR to sustain drug resistance. Thalidomide and As₂O₃ was added to substrate respectively. At the same time, a contrast group was set up. The concentration of VEGF in upper liquid and plasma was examined with enzyme-linked immunosorbent assay （ELISA）and Pgp expression on cell membranes was analyzed by flow cytometry （FCM）.Results : The plasma VEGF concentrations in the patients with AML were significantly higher than the levels of the control （P<0.01）.After added thalidomide and As₂O₃ to substrate, the concentration of VEGF in upper liquid and plasma is significantly lower than the contrast group.After added thalidomide and As₂O₃ to substrate, Pgp expression of the cell of AML and HL60/VCR was not significantly changed.Singnificant correlation was observed between P-gp and VEGF in newly diagnosed AML cases.Conclusions:Leukemia cell can secrete VEGF self;Both thalidomide and As₂O₃ can inhibit the secretion of VEGF of AML and HL-60/VCR;Both thalidomide and As₂O₃ can not affect the Pgp expression on the cell membranes of AML and HL-60/VCR.Test of the two targets together is more effective to the judgment of prognosis in AML.更多还原