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目的 探讨儿童急性淋巴细胞白血病 (ALL)患者在初诊、完全缓解 (CR)及复发不同时期的T细胞受体 (TCR)δV D、γV J基因重排连接区序列的差异及其意义。方法 采用T 载体分子克隆测序、限制性酶切及聚合酶链反应等技术 ,对 34份ALL患者标本进行TCRδ、γ连接区序列动态分析。结果 34份ALL患者标本的TCRδ、γ基因重排序列在初诊、CR及复发不同转归时期显示有差异性和规律性。TCRδV D重排序列分析显示 ,2 4份ALL患者标本中 ,10份初诊患者Vδ2、Dδ3序列较完整 ,其中 7份伴有Dδ3的九碱基序列区T→C突变。 11份CR期患者有明显Vδ2的 3′端、Dδ3的 5′端及七碱基序列重排序列缺失 ,Dδ3九碱基序列区T→C突变消失。初诊与CR期ALL患者Vδ2或Dδ3重排序列缺失率及Dδ3九碱基序列区T→C突变率比较 ,差异均有极显著性意义 (精确概率计算 ,P均 =0 .0 0 1)。 3份复发患者标本均保留Dδ3的 5′端重排序列完整。 10份TCRγV J序列分析结果规律性与δ相似。结论 ALL的TCRδV D、TCRγV J基因重排序列的改变与ALL的发生、疗效及转归有着密切的内在联系。
Objective To investigate the differences of TCRδδV, γVJ gene rearrangement junction sequences in children with acute lymphoblastic leukemia (ALL) at first visit, complete remission (CR) and at different stages of relapse. Methods The TCRδ and γ junctional sequences of 34 ALL patients were analyzed by T vector sequencing, restriction enzyme digestion and polymerase chain reaction (PCR). Results TCRδ and γ gene rearrangements of 34 ALL patients showed no difference and regularity in the newly diagnosed cases and different prognosis of CR and relapse. TCRδV D rearrangement analysis showed that in 24 samples of ALL patients, 10 cases of newly diagnosed patients had complete Vδ2 and Dδ3 sequences, 7 of them were associated with TδC mutation of Dδ3 in the nine base sequence. In 11 CR patients, the 3 ’end of Vδ2 was obvious, the 5’ end of Dδ3 and the rearrangement of the seven base sequence were deleted, and the T → C mutation of the Dδ3 nine base sequence disappeared. The rates of deletion of Vδ2 or Dδ3 rearrangement sequence and T → C mutation rate of Dδ3 nine base sequence region in newly diagnosed and CR stage ALL patients were significantly different (P <0.001). All the 3 recurrent patients retained the 5 ’end rearrangement sequence of Dδ3 intact. The results of 10 TCRγV J sequence analyzes were similar to δ. Conclusion The changes of TCRδV D and TCRγV J gene rearrangements in ALL are closely related to the occurrence, efficacy and outcome of ALL.