Boeckx N, Willemse M J, Szczepanski T, van der Velden V H J, Langerak A W, Vandekerckhove P, van Dongen J J M
Department of Immunology, Erasmus University Rotterdam/University Hospital Rotterdam, Rotterdam, The Netherlands.
Leukemia. 2002 Mar;16(3):368-75. doi: 10.1038/sj.leu.2402387.
PCR-based monitoring of minimal residual disease (MRD) in acute leukemias can be achieved via detection of fusion gene transcripts of chromosome aberrations or detection of immunoglobulin (Ig) and T cell receptor (TCR) gene rearrangements. We wished to assess whether both PCR targets are complementary in acute myeloid leukemia (AML). We investigated 105 consecutive AML cases for the presence of fusion gene transcripts by reverse transcriptase polymerase chain reaction (RT-PCR): AML1-ETO associated with t(8;21), CBFB-MYH11 with inv(16), PML-RARA with t(15;17), BCR-ABL with t(9;22), and MLL-AF4 with t(4;11). In 17 out of 105 AML cases (16%), fusion gene transcripts were found. Ninety-five of these AML patients (13 with fusion gene transcripts) were also investigated for the presence of IGH, IGK, TCRG and TCRD rearrangements by Southern blot and/or PCR heteroduplex analysis and sequencing. In nine out of 95 patients (9.5%), such rearrangements were found. Combined data revealed that only one patient with a fusion gene transcript had a coexistent Ig/TCR rearrangement. The nine AML patients with Ig/TCR rearrangements, as well as five additional AML patients from a previous study were investigated in more detail, revealing that Ig/TCR rearrangements in AML are immature and unusual. The presence of Ig/TCR rearrangements in AML did not correlate with RAG gene expression levels as determined by real-time quantitative PCR. In conclusion, fusion gene transcripts and Ig/TCR rearrangements are infrequent, but complementary MRD-PCR targets in AML.
基于聚合酶链反应（PCR）监测急性白血病微小残留病（MRD），可通过检测染色体畸变的融合基因转录本或免疫球蛋白（Ig）和T细胞受体（TCR）基因重排来实现。我们希望评估这两种PCR靶点在急性髓系白血病（AML）中是否具有互补性。我们通过逆转录聚合酶链反应（RT-PCR）对105例连续的AML病例进行研究，以检测融合基因转录本的存在情况：与t(8;21)相关的AML1-ETO、与inv(16)相关的CBFB-MYH11、与t(15;17)相关的PML-RARA、与t(9;22)相关的BCR-ABL以及与t(4;11)相关的MLL-AF4。在105例AML病例中的17例（16%）发现了融合基因转录本。还通过Southern印迹和/或PCR异源双链分析及测序，对其中95例AML患者（13例有融合基因转录本）进行了IGH、IGK、TCRG和TCRD重排情况的检测。在95例患者中的9例（9.5%）发现了此类重排。综合数据显示，仅有1例有融合基因转录本的患者同时存在Ig/TCR重排。对9例有Ig/TCR重排的AML患者以及之前一项研究中的另外5例AML患者进行了更详细的研究，结果显示AML中的Ig/TCR重排不成熟且不常见。AML中Ig/TCR重排的存在与通过实时定量PCR测定的RAG基因表达水平无关。总之，融合基因转录本和Ig/TCR重排虽不常见，但在AML中是互补的MRD-PCR靶点。
