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目的 :观察 Flt3基因在 AML(急性髓细胞性白血病 )病人骨髓中的表达水平及正常对照骨髓、外周血表达水平的变化及其与病情、疗效的关系。方法 :应用血细胞 Percoll不连续密度梯度分离法分离出骨髓中恶性细胞 ,采用 DIG掺入半定量 RT- PCR,检测 Flt3基因在 2 2例 AML初治病人、5例 AML CR病人的骨髓、10例正常人和非恶性血液病病人 13例外周血、17例骨髓以及 5种白血病细胞株中的表达水平。结果 :5种白血病细胞株中测得 Flt3表达值分别为 NB40 .97,HL- 6 0 1.1,KG- 10 .96 ,K5 6 2及 U937均为 0 ;正常人骨髓为 0 .18～ 0 .45 ,平均 0 .2 2 ;正常人外周血未见表达 ;AML CR骨髓表达为 0 .2～ 0 .6 5 ,平均 0 .2 8。初治 AML 骨髓中 Flt3基因 m RNA表达率为 10 0 % ;其表达水平为 0 .75～ 1.2 4,平均为 0 .96 ,为正常骨髓的 4倍。结论 :在 AML中 Flt3基因为高水平表达 ,主要来源于白血病细胞 ,其检测水平可作为诊断疾病、判断预后的一项灵敏指标。
OBJECTIVE: To observe the expression of Flt3 gene in the bone marrow of AML patients and the expression of Flt3 gene in bone marrow and peripheral blood of normal control and its relationship with disease status and efficacy. Methods: Malignant bone marrow cells were isolated by Percoll discontinuous density gradient method. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) with DIG was used to detect the expression of Flt3 gene in 22 untreated AML patients and 5 AML CR patients. Cases of normal and non-hematological malignancies in 13 cases of peripheral blood, 17 cases of bone marrow and 5 leukemia cell lines in the expression level. Results: The expression of Flt3 in the five leukemia cell lines was NB40.97, HL-60.01, KG-10.96, K562 and U937 respectively. The normal human bone marrow was 0.18-0. 45, an average of 0.22; normal peripheral blood was not expressed; AML CR bone marrow expression was 0.2 ~ 0.56, an average of 0.28. The expression of Flt3 gene m RNA in the naive AML bone marrow was 100%. The expression level of Flt3 mRNA in the untreated AML bone marrow was 0 .75-1.24 with an average of 0.96, which was 4 times of normal bone marrow. Conclusion: Flt3 gene is highly expressed in AML and mainly comes from leukemia cells. The detection level of Flt3 gene can be used as a sensitive indicator to diagnose disease and prognosis.