1 Institute of Hematology, Jinan University , Guangzhou, People's Republic of China .
DNA Cell Biol. 2014 Jan;33(1):49-56. doi: 10.1089/dna.2013.2199. Epub 2013 Dec 13.
Despite significant improvement in our understanding of T-cell acute lymphoblastic leukemia (T-ALL) biology and pathogenesis, many questions remain unanswered. In previous studies, we found a T-ALL case with two malignant T-cell clones with Vδ1Dδ2Dδ3Jδ1 and Vδ2Dδ3Jδ2 rearrangements. In this study, we further characterized T-ALL cases with two malignant clones containing Vδ1Dδ3Jδ1 and Vδ2Dδ1Jδ1 rearrangements using fine-tiling array comparative genomic hybridization, ligation-mediated polymerase chain reaction (LM-PCR), sequencing, and reverse transcription polymerase chain reaction (RT-PCR) analysis. We further analyzed the distribution and clonality of the T-cell receptor (TCR) Vγ and Vδ subfamily T cells in the two T-ALL cases by RT-PCR and GeneScan. Monoclonal Vδ1 and Vδ2 subfamilies were confirmed in both samples, the Vδ3 through Vδ7 subfamilies could not be detected in the T-ALL samples, whereas the oligoclonal Vδ8 subfamily could be identified. Based on the clinical finding that both of the T-ALL cases with two malignant T-cell clones had a poor outcome, we attempted to compare the expression pattern of genes related to T-cell activation and proliferation between cases with the malignant Vδ1 and Vδ2 T-cell clones and T-ALL cases with a mono-malignant Vα T-cell clone. We selected two T-ALL cases with VαJα rearrangements and analyzed the expression level of Notch1, TAL1, and the CARMA-BCL10-MALT-A20-NF-κB pathway genes by real-time PCR. A20 had significantly higher expression in the biclonal compared with the monoclonal T-ALL group (p=0.0354), and there was a trend toward higher expression for the other genes in the biclonal group with the exception of TAL1, although the differences were not statistically significant. In conclusion, we identified two T-ALL cases with biclonal malignant T-cell clones and described the characteristics of the biclonal T-ALL subtype and its gene expression pattern. Thus, our findings may improve the understanding of biclonal T-ALL.
尽管我们对 T 细胞急性淋巴细胞白血病 (T-ALL) 的生物学和发病机制有了显著的了解,但仍有许多问题尚未得到解答。在之前的研究中,我们发现了一例具有两个恶性 T 细胞克隆的 T-ALL 病例,其 Vδ1Dδ2Dδ3Jδ1 和 Vδ2Dδ3Jδ2 重排。在本研究中,我们使用精细平铺阵列比较基因组杂交、连接介导的聚合酶链反应 (LM-PCR)、测序和逆转录聚合酶链反应 (RT-PCR) 分析进一步表征了含有 Vδ1Dδ3Jδ1 和 Vδ2Dδ1Jδ1 重排的两个恶性克隆的 T-ALL 病例。我们还通过 RT-PCR 和 GeneScan 分析进一步分析了两个 T-ALL 病例中 T 细胞受体 (TCR) Vγ和 Vδ亚家族 T 细胞的分布和克隆性。在两个样本中均证实存在单克隆 Vδ1 和 Vδ2 亚家族,而在 T-ALL 样本中无法检测到 Vδ3 至 Vδ7 亚家族,而可以识别寡克隆 Vδ8 亚家族。基于两个 T-ALL 病例均具有两个恶性 T 细胞克隆且预后不良的临床发现,我们试图比较具有恶性 Vδ1 和 Vδ2 T 细胞克隆的病例与具有单恶性 Vα T 细胞克隆的 T-ALL 病例之间与 T 细胞激活和增殖相关的基因的表达模式。我们选择了两个具有 VαJα 重排的 T-ALL 病例,并通过实时 PCR 分析 Notch1、TAL1 和 CARMA-BCL10-MALT-A20-NF-κB 通路基因的表达水平。与单克隆 T-ALL 组相比,双克隆组中 A20 的表达明显更高 (p=0.0354),除 TAL1 外,双克隆组中其他基因的表达也呈上升趋势,尽管差异无统计学意义。总之,我们鉴定了两个具有双克隆恶性 T 细胞克隆的 T-ALL 病例,并描述了双克隆 T-ALL 亚型的特征及其基因表达模式。因此,我们的发现可能有助于提高对双克隆 T-ALL 的认识。