PTK787在SCID小鼠-人AML模型中的抗骨髓血管生成作用研究

The Study of the Mechanism of Treatment with PTK787 for Angiogenesis in the Bone Marrow of SCID Mouse-human Acute Myeloid Leukemia

目的观察酪氨酸激酶抑制剂PTK787在SCID小鼠-人急性髓系白血病模型中对血血管内皮生长因子受体-2mRNA(VEGFR-2mRNA)、外周血CD33阳性表达率以及骨髓微血管密度的影响,探讨PTK787抗急性髓系白血病的作用机制。方法(1)采用RT-PCR法分析30例不同组别之间SCID小鼠VEGFR-2mRNA水平的差异。(2)利用流式细胞仪检测外周血CD33表达。(3)采用SP免疫组化法,用兔抗人vWF(vonWille-brandfactor)多克隆抗体标记骨髓内皮细胞,比较不同组别小鼠之间的骨髓微血管密度(MVD)的差异。结果(1)患病组SCID小鼠外周血VEGFR-2mRNA灰度扫描比为(0.3257±0.0709),高于正常组(0.0826±0.0315)(P<0.05),治疗组为(0.1411±0.0404),与正常组比较(P<0.05)。(2)外周血白细胞CD33的阳性率,治疗组(9.41±1.35%)及患病组(15.08±2.90%)与正常组(0.81±0.11%)比较,P值均<0.01,差异有统计学意义;治疗组与患病组比较,P<0.05,差异有统计学意义。(3)患病组SCID小鼠中骨髓微血管数为(20.4±3.2)个/HP,显著高于正常组(14.2±2.1)个/HP(P<0.001),治疗组为(15.6±2.4)个/HP,与正常组相似(P>0.05)。结论PTK787在体内可以有效的抑制血管生成,一定程度具有抗白血病细胞增生的作用。

Objective To observe the effects of the tyrosine kinase inhibitor PTK787 on the expression of VEGFR - 2mRNA, the CD33 positive proportion of peripheral blood cells and bone marrow microvascular density in SCID mouse - human acute myeloid leukemia （AML） , and explore the role of PTK787 on anti - acute myeloid leukemia. Methods （ 1 ） The expression of vascular endothelial growth factor receptor - 2 mRNA （ VEGFR - 2 mRNA ） is detected by reverse transcription - polymerase chain reaction. （2） CD33 of peripheral white blood ceils surface of all mice were assayed by flowing cytometry （ FCM ）. （ 3 ） Immunohistochemical technique（SP） was employed to stain microvascular endothelial cells in each bone marrow specimen section with anti - human yon Willbrand factor. The diversity of bone marrow microvascular density among different groups of SCID mouse is to determine. Results 1. Compare with the disease group （0. 3257 ± 0. 0709） , VEGFR - 2 mRNA in freating group （0. 1411 ± 0. 0404） was decreased obviously; Difference between the normal group（0. 0826 ± 0. 0315） and the treating group on the expression of VEGFR -2 mRNA had statistic significant. 2. The CD33 positive proportion of peripheral blood cells, the difference between other groups and normal group （0.81 ± 0.11% ）are significant in statistics, P 〈 0.01 ;the difference between PTK787 treating group（ 9.41 ± 1.35 % ） and disease group （ 15.08 ± 2.90% ） is significant in statistics, P 〈 0.05; 3. The bone marrow microvessel density（MVD） in the disease group[ （20.4 ± 3.2）/high -power field（HP） ] was significantly higher than that in the control group[ （ 14.2 ± 2. 1 ）/HP, P 〈 0. 001 ]. No significant difference was found between the treating group [ （ 15.6 ± 2.4）/HP] and the control group（ P 〉 0.05 ）. Conclusion The bone marrow microvascular density in disease group was higher than control. PTK787 has antiangiogenesis role in acute leukemia.