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目的测定cAMP反应元件结合蛋白(Cyclic-AMP Response Binding Protein,CREB)在不同类型儿童急性髓细胞白血病(AML)中的表达,探讨其在儿童AML发生、发展和预后判断中的意义。方法(1)用实时定量RT-PCR法检测不同类型AML患儿骨髓单个核细胞内CREB的mR-NA水平的表达,以非白血病患儿(NL)作对照。(2)用ELISA法测定AML患儿骨髓单个核细胞内CREB总蛋白水平和蛋白磷酸化水平的表达。(3)AML患儿中,是否存在MICM分型的高危因素与CREB的mRNA是否高表达相比较。(4)对3例AML患儿骨髓单个核细胞内CREB的mRNA水平进行动态观察。结果(1)NL、低危型AML(LR-AML)、高危型AML(HR-AML)各组患儿骨髓单个核细胞内CREB的mRNA水平的表达量分别为1.13±0.81、1.23±1.14、11.88±19.64;其中,LR-AML组与NL组无明显差别,HR-AML组明显高于NL组、LR-AML组,差别有统计学意义,P<0.05。(2)HR-AML组患儿骨髓单个核细胞内CREB总蛋白水平的表达(ng/106cell)明显高于LR-AML组,分别为9.97±13.84、1.06±1.14,P=0.005;HR-AML组患儿骨髓单个核细胞内CREB蛋白磷酸化水平的表达(U/106cell)明显高于LR-AML组,分别为19.76±29.11、1.36±1.84,P=0.01。(3)在AML患儿中,CREB的mRNA水平、总蛋白水平、蛋白磷酸化水平的表达量间两两存在正相关关系。(4)HR-AML患儿中,60.00%的患儿存在MICM分型的高危因素,70.00%的患儿CREB的mRNA呈高表达,两者相结合诊断符合率可达90.00%。(5)CREB的mRNA表达量的升高提示AML的未缓解。结论(1)CREB在儿童HR-AML中呈高表达。(2)儿童AML中CREB的mRNA水平、总蛋白水平、蛋白磷酸化水平的表达量间存在正相关关系。(3)CREB的表达与MICM分型从不同的水平判断AML患儿的预后,对临床危险度分型有一定的意义。(4)实时定量RT-PCR进行CREB的mRNA表达量的动态监测对白血病疗效的判断意义值得进一步探讨。
Objective To determine the expression of cAMP response element binding protein (CREB) in children with acute myeloid leukemia (AML) and explore its significance in the development, prognosis and prognosis of childhood AML. Methods (1) The expression of mR-NA of CREB in bone marrow mononuclear cells of children with different types of AML was detected by real-time quantitative RT-PCR. Non-leukemia children (NL) were used as controls. (2) The expression of CREB total protein and protein phosphorylation in bone marrow mononuclear cells of children with AML were measured by ELISA. (3) In children with AML, is there a high risk of MICM typing compared with the high expression of CREB mRNA? (4) Dynamic observation of CREB mRNA level in bone marrow mononuclear cells of 3 children with AML. Results (1) The expression levels of CREB mRNA in bone marrow mononuclear cells of NL, low-risk AML (LR-AML) and high-risk AML (HR-AML) groups were 1.13 ± 0.81 and 1.23 ± 1.14, 11.88 ± 19.64. There was no significant difference between LR-AML group and NL group, HR-AML group was significantly higher than NL group and LR-AML group, the difference was statistically significant, P <0.05. (2) The expression of CREB protein in bone marrow mononuclear cells of children with HR-AML (ng / 106cell) was significantly higher than LR-AML group (9.97 ± 13.84, 1.06 ± 1.14, P = 0.005) The expression of CREB protein phosphorylation in bone marrow mononuclear cells (U / 106cell) was significantly higher than LR-AML group (19.76 ± 29.11,1.36 ± 1.84, P = 0.01). (3) There was a positive correlation between CREB mRNA level, total protein level and protein phosphorylation in children with AML. (4) Among children with HR-AML, 60.00% of children had risk factors for MICM classification, and 70.00% of children had high expression of CREB mRNA. The coincidence rate of the two was 90.00%. (5) The increase of mRNA expression of CREB suggests that AML is not relieved. Conclusions (1) CREB is highly expressed in children with HR-AML. (2) There was a positive correlation between CREB mRNA level, total protein level and protein phosphorylation in children with AML. (3) The expression of CREB and MICM classification to judge the prognosis of children with AML from different levels have certain significance to the classification of clinical risk. (4) Real-time quantitative RT-PCR to detect CREB mRNA expression in the dynamic monitoring of the significance of the judgment of the efficacy of leukemia should be further explored.