高白细胞性急性髓系白血病细胞基因表达谱的研究

Gene Miaroarray Profile Study of Leukemia Blasts in Hyperleukocytic Acute Myeloid Leukemia

[目的]综合分析高白细胞性急性髓系白血病不同预后患者祖细胞集落形成及对化疗药物的敏感性以及差异表达的基因。[方法]半固体集落形成法观察不同预后患者骨髓祖细胞集落形成及对化疗药物的敏感性。采用Affymetrix公司人类寡核苷酸表达谱芯片检测患者初诊时骨髓单个核细胞基因表达谱,筛选出差异表达数据。[结果](1)患者骨髓祖细胞集落形成和化疗药物的敏感性与预后密切相关,未缓解者的骨髓祖细胞集落明显高于完全缓解患者,且对化疗药物不敏感。(2)完全缓解患者白血病细胞表达上调2倍以上的基因109条,占0.5%。下调2倍以上的基因134条,占0.6%,其中与预后密切相关的为TRIM16、TM4SF1。CD34和DNMT3B。(3)差异表达的基因按功能分成离子通道运输、细胞周期、细胞骨架运动、细胞凋亡等14类。上调的主要为DNA合成修复、细胞凋亡、细胞周期和应激相关等4类;而下调的主要为蛋白翻译合成、信号传导传递、发育相关和细胞骨架运动等4类。[结论]特定的基因表达模式决定患者预后。

[Objective] In order to explore pathogenesis and prognosis of gene expression in hyperleukocytic AML,we analyze the proliferation potency,chemotherapeutic drug susceptibility of leukemic progenitor cells and gene expression profiles by oligonucleotide microarray.[Method]The proliferation potency and chemotherapeutic drug susceptibility of leukemic progenitor cells were detected by colony formation unite assay of bone marrow from untreated hyperleukocytic AML patients with diverse prognosis.The gene expression profiles of leukemia blasts were analyzed from peripheral blood of these untreated patients with the oligonucleotide microarray from Affymetrix Gene Expression Service Lab.The diversity of gene expression was screened and analyzed from hybridization of cDNA chip.[Results](1)The colony formation of 105 cells from bone marrow was 89,95 respectively from two CR cases,and 191,187 separately from two NR patients.The suppression rates of chemotherapeutic drug were higher than 30% in CR cases except that 3 cases to Adr,and lower than 30% in NR patients except that 2 cases to VP16,suggesting that leukemic progenitor cells of NR patients had higher proliferation potency and lower susceptibility to cytotoxic drug.(2)The gene expression level in two CR cases was identified as 109 pieces of up regulation gene and 134 pieces of down regulation gene compared with two NR cases.TRIM16 and TM4SF1,CD34 and DNMT3B among above diversity genes were closely interrelated with prognosis as acute leukemia-specific genes.(3)Being compared CR with NR of hyperleukocytic AML,both up-regulated and down-regulated genes by more than two fold could be classified as fourteen sorts according to their function,which had ion channel transport,cell cycle,cytoskeleton action,apoptosis,stress reaction,DNA synthesis/repair,transcription regulation,receptor,immune regulation,intracellular signal pathway,protein translation/synthesis,metabolism,development and other unclassified functions.(4)The four kinds of genes related with DNA synthesis/repair,apoptosis,cell cycle and stress reaction were differential expression of up regulation,while the other four kinds of protein translation/synthesis,intracellular signal pathway,development genes and cytoskeleton action were differential expression of down regulation in CR patients.[Conclusion]The designated gene expression profiles determine the prognosis of hyperleukocytic AML.This significant gene expression profiles may provide useful evidence for the pathogenesis and prognosis in high dangerous hyperleukocytic AML.