| ObjectiveBy detecting the expression of PRAME in bone marrow (BM) / peripheral blood (PB) mononuclear cells of children with acute leukemia ,and compared with the expression of WT1 gene. To Explore the relationship of PRAME gene and WT1 gene, as well as with the curative effect and the prognosis. Providing a feasible method and a molecular Marker for the realization of monitoring minimal residual diseaseMethodsThe expression of PRAME gene and expression level was measured by semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR) in the MB and PB samples of 51 children with acute leukemia. Non-remission group (relapse +newly diagnosed) is 40 cases, including 34 cases of newly diagnosed (ALL 24cases, AML 10cases); relapse in 6 cases (ALL 5cases, AML 1cases).The remission group is 11 children, (ALL 9cases, AML 2cases). The control group is 25 cases. In the same period,WT1 mRNA expression was measured, combined with efficacy, prognosis, a comprehensive analysis.4 cases of children with newly diagnosed AL tracking follow-up, compared detection of bone marrow PRAME mRNA, WT1 mRNA expression and the expression level .To analysis of gene expression changes with disease progression. To clear the application of RT-PCR technique used in detection of PRAME gene expression is feasible.Results1. Overexpression of PRAME mRNA was found in Non-remission group is 17, the total positive rate was 42.5%. In newly diagnosed group and relapse group , PRAME mRNA expression rate were 35.3%,50.0%, the two groups of comparisons has not statistically significant difference (p= 0.195). In ALL group and AML group, PRAME mRNA expression rate are 41.4%,45.5%, there are no statistically significant difference (p= 1.000).The Non-remission Group is compared with the remission group (p1 = 0.030)and the control group (p2 = 0.000) ,there are no statistically significant difference.2. In the Non-remission group,the expression of PRAME gene and WT1 gene in the same rate (both positive or negative)are 57.5% (23 cases).Only one of gene expression positive rate was 67.5% (27 cases).In children with AL,the expression of PRAME and WT1 have not Statistical significant correlation (p = 0.337).3. By using semi-quantitative detection,we found that PRAME mRNA, WT1 mRNA expression level with no statistical significant differences between MB and MB ,which were collected in 6 newly diagnosied children.4. 34 cases of newly diagnosed children with CR rate 88.2% (30 cases). Which the expression of PRAME gene,WT1 gene were both positive and negative groups of children with CR rates were 62.5%, 100%, the two groups of comparisons no statistically significant difference (p = 0.049).5. Short-term follow-up (1 month～13 months) observed four cases of children with newly diagnosed AL. Of which 2 cases were achieved complete remission, the expression of PRAME gene and WT1 gene decreased and gradually turned negative, is still under follow-up, PRAME gene and WT1 gene expression continued negative, children were clinical complete remission. Another two cases below the 1st course of treatment to alleviate, PRAME gene, WT1 gene continuing high levels of expression, giving up treatment.Conclusion1.PRAME gene in children with AL showed a high level ,can be expressed as an important molecular marker.2.The expression level of Bone marrow and peripheral blood is consistent,can be applied to replace theBM to PB for monitoring MRD.3.PRAME gene and WT1 gene expression in the AL in children was no significant correlation, but the both positive expression was significantly reduced complete remission rate. Combination of the two used in the monitoring of MRD can be found monitoring the vast majority of AL children.4.Using RT-PCR, track and monitor the changes of expression levels of PRAME mRNA can be a certain extent to understand the load in children with AL, help to guide treatment, is expected to be indicators for monitoring of minimal residual disease. |
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