兒童急性白血病Notch1、Jagged1基因和蛋白表達及臨床意義
發(fā)布時間：2019-06-19 10:05
【摘要】：目的 通過應(yīng)用RT-PCR與免疫細胞化學（ICC）技術(shù)檢測急性白血病(AL)兒童骨髓(MB)或外周血(BP)細胞中Notch相關(guān)分子mRNA及Notch1蛋白的表達情況，探討Notch信號通路在兒童各型急性白血病發(fā)生中的致病機制，為兒童AL的靶向治療提供新的理論依據(jù)。 方法 第一部分通過應(yīng)用RT-PCR技術(shù)檢測AL兒童MB或BP單個核細胞中Notch信號的Notch1受體與Jagged1配體以及內(nèi)參照β-肌動蛋白(β-actin)的mRNA的表達。47例初診急性白血病兒童（其中B-ALL26例，T-ALL6例和AML15例）和同期非惡性血液病BM標本5例；門診體檢正常兒童PB標本15例，評價Notch1和Jagged1基因陽性表達率；通過Notch1基因=Notch1基因表達/β-actin表達和Jagged1基因=Jagged1基因表達/β-actin表達，計算檢測基因的表達水平。 第二部分通過應(yīng)用免疫細胞化學（ICC）技術(shù)檢測急性白血病兒童骨髓涂片細胞中Notch1蛋白表達。42例初診急性白血病兒童（其中B-ALL25例，T-ALL7例和AML10例）骨髓涂片和同期非惡性血液病BM涂片標本22例作為對照，每例高倍鏡視野下隨機計數(shù)100個細胞，從Notch1蛋白陽性表達細胞數(shù)和細胞著色程度進行分析。 結(jié)果 第一部分Notch1基因的陽性率：ALL及AML組顯著高于對照組，差異有統(tǒng)計學意義（P0.05）；ALL組與AML組比較（P0.05），其差異無統(tǒng)計學意義。Notch1基因的表達水平：ALL組及AML組和對照組比較，，ALL組與AML初發(fā)組比較，差異無統(tǒng)計學意義（P0.05）。 T-ALL組與B-ALL Notch1基因陽性率差異無統(tǒng)計學意義（P0.05）；但是，兩組Notch1基因表達水平差異有統(tǒng)計學意義（t=3.835，P=0.001）。Jagged1基因的陽性率：ALL組與對照組比較（P0.05）、AML組與對照組比較及ALL組與AML組比較（P0.05）差異均無統(tǒng)計學意義。Jagged1基因的表達水平：ALL組及AML組與對照組比較，差異有統(tǒng)計學意義(P0.05)。ALL組與AML組比較，其差異無統(tǒng)計學意義（P0.05）。T-ALL組與B-ALL在基因表達陽性率及基因表達水平兩方面差異無統(tǒng)計學意義（P0.05）。 發(fā)病初期同時收集BM及PB的12例AL患兒，進行BM和PB中基因表達水平的半定量比較，發(fā)現(xiàn)Notch1基因、Jagged1基因在BM及PB中表達一致，經(jīng)統(tǒng)計分析，其差異無統(tǒng)計學意義（P0.05）。第二部分Notch1受體蛋白的陽性細胞表達率：T-ALL組及AML組與對照組相比，差異有統(tǒng)計學意義（P0.05），B-ALL與對照組相比差異無統(tǒng)計學意義（P0.05），ALL組與對照組相比，差異無統(tǒng)計學意義（P0.05）。T-ALL患兒和B-ALL患兒進行比較，差異有統(tǒng)計學意義（P0.05）。 Notch1蛋白的陽性細胞表達率依照ALL-LR、ALL-MR、ALL-HR的次序升高，HR組高于LR組及MR組，但組間比較，差異均無統(tǒng)計學意義（P0.05）。 結(jié)論 1.在不同類型兒童ALL中，Notch1的表達有明顯差異，T-ALL患兒中的Notch1表達明顯高于B-ALL。 2.Notch1在兒童AML異常表達，提示Notch1可能在兒童AML中起著重要作用。 3.Jagged1在ALL及AML患兒中異常表達，Jagged1可能在兒童AL中也起著重要作用。 4.骨髓及外周血中Notch1mRNA、Jagged1mRNA表達水平一致，可用外周血替代骨髓進行Notch1、Jagged1基因mRNA檢測。 5.Notch1的mRNA及蛋白表達異常對于兒童急性白血病免疫分型有一定指導意義。
[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
【學位授予單位】：第四軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R733.71
本文編號：2502269
[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
【學位授予單位】：第四軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R733.71
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