儿童急性B淋巴细胞白血病APRIL及其受体mRNA水平的定量测定

目的:采用实时荧光定量PCR(RFQ-PCR)技术分析儿童急性B淋巴细胞白血病患儿外周血APRIL及其受体mRNA准确含量,并以靶基因和内参β2MmRNA含量的比值作为评价靶基因表达水平的指标。方法:在APRIL及其受体基因的高保守区设计相应引物和荧光探针,实时检测PCR产物的荧光强度,根据标准品建立的标准曲线,由软件自动计算出待测样本中靶基因中APRIL和BCMA的水平。结果:18例急性B淋巴细胞白血病样本的APRIL及其受体BCMA和TACImRNA水平的表达水平均显著高于正常儿童(P<0.05)。结论:发现APRIL在儿童急性B淋巴细胞白血病的发病过程中可能起了重要作用,并通过受体BCMA和TACI发挥作用。

Measurement of the expression level of APRIL and its receptors in B-ALL by real-time fluorenscence quantitative method

Objective:To establish real-time fluorescence quantitative polymerase chain reaction(RFQ-PCR) for measurement of APRIL expression level and its receptors in B-ALL children.Methods:Specific primers and TaqMan probes of APRIL and its receptors were designed,and fluorescence of the PCR products were detected continuously during amplification.According to the standard curves created by plasmid DNA,the expression level of target genes in clinical samples were determined by using software,and the results were presented as the ratios of target genes' mRNA to β2 Microgluobulin(β2M)'s mRNA.Results:The results from 18 samples showed that the expression level of APRIL,BCMA and TACI in the peripheral blood of B-ALL were significantly higher than that in normal children(P<0.05).Conclusions:APRIL may play a great role in the development,and progress of B-ALL through its receptor B cell maturation antigen(BCMA) and transmembrane activator and CAML interactor(TACI).