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目的通过检测急性白血病(AL)患儿维生素D受体(VDR)mRNA和蛋白质表达,从转录和翻译两个水平探讨VDR在AL中的表达和意义。方法收集2009年1月至12月广西医科大学第一附属医院儿童血液科住院或门诊未经治疗的30例急性淋巴细胞白血病(ALL)患儿(ALL1组)、12例急性髓细胞白血病(AML)患儿(AML1组)、30例非肿瘤性血液病患儿(对照1组)的骨髓标本,用实时荧光定量聚合酶链反应(PCR)方法检测3组骨髓细胞VDRmRNA表达。收集2009年9月至2010年4月广西医科大学第一附属医院儿童血液科住院或门诊未经治疗的30例ALL患儿(ALL2组)、10例AML患儿(AML2组)和30名健康体检儿童(对照2组)的外周血标本,提取外周血单个核细胞(PBMC),应用免疫细胞化学法和蛋白免疫印迹法检测VDR蛋白表达。结果 ALL1组、AML1组和对照1组骨髓均有VDRmRNA表达。ALL1组VDRmRNA表达量为(1.06±0.31)、AML1组VDRmRNA表达量为(1.13±0.34),均明显低于对照1组(3.10±0.18),3组差异有统计学意义(F=1701.00,P<0.01);而ALL1组与AML1组间VDRmRNA表达差异无统计学意义(P>0.05)。免疫细胞化学法显示,ALL2组外周血VDR蛋白表达(27.82±1.78)%、AML2组外周血VDR蛋白表达(27.10±2.44)%,明显低于对照2组(59.02±3.46)%,3组间差异有统计学意义(F=1150.26,P<0.01);而ALL2组与AML2组VDR蛋白表达差异无统计学意义(P>0.05)。蛋白免疫印迹法显示,ALL2组VDR蛋白表达(0.299±0.071)、AML2组VDR蛋白表达(0.290±0.094),明显低于对照2组(0.710±0.041),3组间差异有统计学意义(F=356.434,P<0.01);而ALL2组与AML2组VDR蛋白表达差异无统计学意义(P>0.05)。结论所有检测的AL患儿骨髓及外周血单个核细胞中均表达VDR,但VDR表达可能与AL分类无关。
Objective To investigate the expression and significance of VDR in AL from both transcriptional and translational levels by detecting the expression of vitamin D receptor (VDR) mRNA and protein in children with acute leukemia (AL). Methods Thirty patients with acute lymphoblastic leukemia (ALL) (ALL1 group) and 12 patients with acute myeloid leukemia (AML) were admitted to the First Affiliated Hospital of Guangxi Medical University from January 2009 to December 2005. ) Patients (AML1 group) and 30 non-neoplastic hematologic patients (control group 1). The expression of VDR mRNA in three groups of bone marrow cells was detected by real-time fluorescence quantitative polymerase chain reaction (PCR) A total of 30 ALL children (ALL2 group), 10 AML children (AML2 group) and 30 healthy children (AML2 group) were enrolled from September 2009 to April 2010 in the First Affiliated Hospital of Guangxi Medical University. Peripheral blood mononuclear cells (PBMCs) were extracted from peripheral blood of healthy children (control group 2). The expression of VDR protein was detected by immunocytochemistry and Western blot. Results The bone marrow of ALL1 group, AML1 group and control group 1 had VDR mRNA expression. The expression of VDR mRNA in ALL1 group was (1.06 ± 0.31) and that in AML1 group was (1.13 ± 0.34), which were significantly lower than that in control group 1 (3.10 ± 0.18), the difference was statistically significant (F = 1701.00, P <0.01). There was no significant difference in VDR mRNA expression between ALL1 group and AML1 group (P> 0.05). Immunocytochemistry showed that the expression of VDR protein in peripheral blood of ALL2 group was (27.82 ± 1.78)%, that of AML2 group was (27.10 ± 2.44)%, significantly lower than that of control group 2 (59.02 ± 3.46)%, The difference was statistically significant (F = 1150.26, P <0.01). There was no significant difference in VDR protein expression between ALL2 group and AML2 group (P> 0.05). Western blotting showed that VDR protein expression in ALL2 group (0.299 ± 0.071) and VDR protein expression in AML2 group (0.290 ± 0.094) was significantly lower than that in control group 2 (0.710 ± 0.041), with significant difference between the three groups = 356.434, P <0.01). There was no significant difference in VDR protein expression between ALL2 group and AML2 group (P> 0.05). Conclusions VDR is expressed in all myeloid and peripheral blood mononuclear cells in all AL patients, but VDR expression may not be related to AL classification.