Detection of WT1 gene in acute myeloid leukemia children by real-time fluorescent quantitative RT-PCR and its clinical significance / 中国实验血液学杂志

ABSTRACT

Objective of this study was to establish a SYBR Green Ireal-time reverse transcription-polymerase chain reaction (RT-PCR) for quantitative detection of WT1 gene mRNA in children with acute myeloid leukemia (AML) and investigate its clinical significance. SYBR Green Ireal-time RT-PCR was used to quantitatively detect the mRNA expression of WT1 gene in 30 newly diagnosed AML patients, 12 cases of remission (30), 18 relapsed patients and 30 cases of normal bone marrow cell morphology, and dynamically to detect the expression of WT1 gene in 20 newly diagnosed AML children. ABL served as internal reference gene, and the 2(-&Delta;&Delta;ct) method was used to calculate the relative expression. The results showed that (1) the expression of WT1 gene in newly diagnosed AML children was higher than that of the normal controls and the patients with remission (p < 0.001); there were no significant difference of WT1 gene expression between AML patients with remission and normal controls (p > 0.05), which were same as in relapsed patients and newly diagnosed patients (p > 0.05); (2) WT1 gene in 20 newly diagnosed AML children highly expressed before the children were initially treated, decreased when they were complete remission, then expression increased again when their AML relapsed. The WT1 gene expression level began to rise in 5 cases before clinical relapse at 5 - 7 months; (3) the complete remission rate (CR) and 3 year overall survival (OS) did not show significant difference between the WT1-positive group and negative group when dynamically monitoring WT1 gene expression of 20 newly diagnosed children with AML. 3-year OS of WT1-positive group at the 22 - 30 days after initial treatment was significantly lower than that of the negative group (p < 0.05). It is concluded that SYBR Green Ireal-time RT-PCR is a rapid, efficient, sensitive and specific method. WT1 gene in AML childhood plays a role of cancer-promoting. The change of WT1 gene expression level contributes to evaluate the therapeutic efficacy, detect the minimal residual diseases and analyze the prognosis.