儿童急性白血病Notch1、Jagged1基因和蛋白表达及临床意义

发布时间：2019-06-19 10:05

【摘要】：目的 通过应用RT-PCR与免疫细胞化学（ICC）技术检测急性白血病(AL)儿童骨髓(MB)或外周血(BP)细胞中Notch相关分子mRNA及Notch1蛋白的表达情况，探讨Notch信号通路在儿童各型急性白血病发生中的致病机制，为儿童AL的靶向治疗提供新的理论依据。 方法 第一部分通过应用RT-PCR技术检测AL儿童MB或BP单个核细胞中Notch信号的Notch1受体与Jagged1配体以及内参照β-肌动蛋白(β-actin)的mRNA的表达。47例初诊急性白血病儿童（其中B-ALL26例，T-ALL6例和AML15例）和同期非恶性血液病BM标本5例；门诊体检正常儿童PB标本15例，评价Notch1和Jagged1基因阳性表达率；通过Notch1基因=Notch1基因表达/β-actin表达和Jagged1基因=Jagged1基因表达/β-actin表达，计算检测基因的表达水平。 第二部分通过应用免疫细胞化学（ICC）技术检测急性白血病儿童骨髓涂片细胞中Notch1蛋白表达。42例初诊急性白血病儿童（其中B-ALL25例，T-ALL7例和AML10例）骨髓涂片和同期非恶性血液病BM涂片标本22例作为对照，每例高倍镜视野下随机计数100个细胞，从Notch1蛋白阳性表达细胞数和细胞着色程度进行分析。 结果 第一部分Notch1基因的阳性率：ALL及AML组显著高于对照组，差异有统计学意义（P0.05）；ALL组与AML组比较（P0.05），其差异无统计学意义。Notch1基因的表达水平：ALL组及AML组和对照组比较，，ALL组与AML初发组比较，差异无统计学意义（P0.05）。 T-ALL组与B-ALL Notch1基因阳性率差异无统计学意义（P0.05）；但是，两组Notch1基因表达水平差异有统计学意义（t=3.835，P=0.001）。Jagged1基因的阳性率：ALL组与对照组比较（P0.05）、AML组与对照组比较及ALL组与AML组比较（P0.05）差异均无统计学意义。Jagged1基因的表达水平：ALL组及AML组与对照组比较，差异有统计学意义(P0.05)。ALL组与AML组比较，其差异无统计学意义（P0.05）。T-ALL组与B-ALL在基因表达阳性率及基因表达水平两方面差异无统计学意义（P0.05）。 发病初期同时收集BM及PB的12例AL患儿，进行BM和PB中基因表达水平的半定量比较，发现Notch1基因、Jagged1基因在BM及PB中表达一致，经统计分析，其差异无统计学意义（P0.05）。第二部分Notch1受体蛋白的阳性细胞表达率：T-ALL组及AML组与对照组相比，差异有统计学意义（P0.05），B-ALL与对照组相比差异无统计学意义（P0.05），ALL组与对照组相比，差异无统计学意义（P0.05）。T-ALL患儿和B-ALL患儿进行比较，差异有统计学意义（P0.05）。 Notch1蛋白的阳性细胞表达率依照ALL-LR、ALL-MR、ALL-HR的次序升高，HR组高于LR组及MR组，但组间比较，差异均无统计学意义（P0.05）。 结论 1.在不同类型儿童ALL中，Notch1的表达有明显差异，T-ALL患儿中的Notch1表达明显高于B-ALL。 2.Notch1在儿童AML异常表达，提示Notch1可能在儿童AML中起着重要作用。 3.Jagged1在ALL及AML患儿中异常表达，Jagged1可能在儿童AL中也起着重要作用。 4.骨髓及外周血中Notch1mRNA、Jagged1mRNA表达水平一致，可用外周血替代骨髓进行Notch1、Jagged1基因mRNA检测。 5.Notch1的mRNA及蛋白表达异常对于儿童急性白血病免疫分型有一定指导意义。
[Abstract]:Purpose To study the expression of Notch-related molecular mRNA and Notch1 protein in the bone marrow (MB) or peripheral blood (BP) cells of children with acute leukemia (AL) by RT-PCR and immunocytochemistry (ICC), and to study the pathogenesis of Notch signaling pathway in children with acute leukemia. The system provides a new theory for the targeted therapy of AL. It was reported. Methods The first part of the method was used to detect the expression of the Notch1 receptor and the Jagged1 ligand of the Notch signal in a single core cell of AL or BP by RT-PCR. The positive expression rate of Notch1 and Jagged1 gene was evaluated in 15 of the LL26, T-ALL6 and AML15 cases, and the positive expression rate of the Notch1 and Jagged1 genes in 15 cases of normal children with normal physical examination; the expression of the Notch1 gene, the Notch1 gene and the Jagged1 gene, the expression of the Jagged1 gene, and the expression of the Jagged1 gene/ HCO3-ac the expression of tin, the calculation of the detection gene, The expression level of Notch1 in the bone marrow smear cells of children with acute leukemia was detected by using the immunocytochemistry (ICC) technique.42 cases of acute leukemia (B-ALL25, T-ALL7 and AML10) were detected in the bone marrow smear and the non-malignant hemopathy BM-smear specimen in the same period. Twenty-two cases were used as control,100 cells were randomly counted from the visual field of each high-power mirror, and the number of cells and the number of cells were expressed from the Notch1 protein. color range Results The positive rate of Notch1 gene in the first part was significantly higher than that of the control group (P0.05). There was no significant difference in the expression level of the Notch1 gene. The expression level of the Notch1 gene was compared with that of the ALL group and the AML group and the control group. The difference of the positive rate of Notch1 gene in the T-ALL group and the B-ALL gene was not statistically significant (P0.05); however, there was a significant difference in the expression level of the Notch1 gene in the two groups (t = 3). .835, P = 0.001). The positive rate of Jagged1 gene: the positive rate of the Jagged1 gene: the comparison between the ALL group and the control group (P0.05), the comparison between the AML group and the control group and the group of the ALL group and the AML group (P0 There was no significant difference in the expression level of the Jagged1 gene. The difference between the ALL group and the AML group was not significant (P0.05). The difference between the ALL group and the AML group was not significant (P0.05). The difference between the positive rate of the gene expression and the expression level of the gene in the T-ALL group and the B-ALL group was not significant (P0.05). There was no statistical significance (P0.05). In the early stage,12 AL children with BM and PB were collected, and the expression level of the gene in BM and PB was compared. The Notch1 gene and Jagged1 gene were found to be consistent in BM and PB. There was no significant difference in the expression of the positive cells of the second part of the Notch1 receptor (P0.05). The difference between the T-ALL group and the control group was statistically significant (P0.05), and the difference of the B-ALL and the control group was not significant (P0.05). (05) There was no significant difference between the ALL group and the control group (P0.05). The children with T-ALL and B-ALL were compared. The expression rate of the positive cells of the Notch1 protein was higher according to the order of ALL-LR, ALL-MR and ALL-HR, and the HR group was higher than that of the LR group and the MR group. , poor There was no significant difference in the expression of Notch1 in ALL of different types of ALL, T-ALL The expression of Notch1 in the children was significantly higher than that of B-ALL. note that Notch1 may play an important role in children's AML.3. Jagged1 is abnormal in ALL and AML children The expression of Notch1mRNA and Jagged1mRNA in bone marrow and peripheral blood is consistent with the expression of Notch1mRNA and Jagged1mRNA in bone marrow and peripheral blood. Notch1, Jagged1 gene mRNA detection in the generation of bone marrow.5. mR of Notch1
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R733.71

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