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沙利度胺联合三氧化二砷对急性髓系白血病细胞表达VEGF和P-gp的影响

Study on Effect of Combination of Thalidomide and As2O3 on VEGF and P-gp Expression in AML

【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。

【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.

【关键词】 VEGFP-gP沙利度胺As2O3
【Key words】 vascular endothelial growth factorPgpthalidomideAs2O3
  • 【网络出版投稿人】 南昌大学
  • 【网络出版年期】2009年 01期
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