VEGF和P-gp在急性髓系白血病中的表达及意义

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P-糖蛋白（P-gp）的影响。方法收集22例AML患者[完全缓解（CR）患者15例（CR组）和未缓解患者7例（非CR组）]的骨髓液,用Ficoll淋巴细胞分离液分离出骨髓单个核细胞（BMMNC）,按实验设计分为a、d、f、h组：a组加BMMNC 800μL;d组加BMMNC 800μL＋TLD 100μL（TLD终质量浓度300μg.mL-1）;f组加BMMNC 800μL＋As2O3100μL（As2O3终浓度25μmol.L-1）;h组加BMMNC 800μL＋TLD 100μL＋As2O3100μL（TLD终质量浓度300μg.mL-1,As2O3终浓度25μmol.L-1）。均于37℃、5%CO2、饱和湿度条件下传代培养,取对数生长期细胞,各组均培养72 h。用双抗体夹心酶联免疫法（ELISA）法检测细胞培养上清液中VEGF的含量,用流式细胞术（FCM）检测细胞膜上MDR1/P-gp的表达率。结果非CR组细胞培养上清中的VEGF含量、细胞膜上P-gp的表达水平均显著高于CR组（P〈0.01）;非CR患者P-gp表达率与VEGF含量具有一定的正相关性[r=0.646（a组）,r=0.695（d组）,r=0.645（f组）,r=0.650（h组）;均P〈0.05]。结论对于初治AML患者,化疗前后VEGF的含量与细胞膜表面P-gp表达的联合检测可作为监测病情和判断预后的指标。

Expression and Significance of VEGF and P-gp in Acute Myeloid Leukemia

Objective To investigate the effects of thalidomide and As2O3 on vascular endothelial growth factor（VEGF） secretion and P-glycoprotein（P-gp） expression in acute myeloid leukemia（AML） cells.Methods Bone marrow mononuclear cells（BMMNC） were isolated from marrow aspirates by using Ficoll-isopaque solution in 15 complete remission（CR） patients and 7 non-complete remission（NCR） patients.Cells in logarithmic growth phase were chosen and cultured at 37 ℃ in a 5% CO2,water-saturated atmosphere for 72 hours.BMMNC were treated with 300 μg·mL-1 thalidomide（group d） or 25 μmol·L-1 As2O3（group f） alone or in combination（group h）.The control cells（group a） received the medium only.The concentration of VEGF in culture supernatants was examined by double antibody sandwich enzyme-linked immunosorbent assay（DAS-ELISA）.The expression of P-gp in cell membranes was detected by flow cytometry（FCM）.Results Compared with CR patients,VEGF concentration and P-gp expression significantly increased in NCR patients.There was a correlation between P-gp expression and VEGF concentration in NCR patients（group a,r=0.646;group d,r=0.695;group f,r=0.645;group h,r=0.650;all P0.05）.Conclusion The combined detection of P-gp expression and VEGF secretion may be useful for monitoring of disease progression and prognosis in AML.