儿童急性淋巴细胞白血病危险度相关基因表达谱的初步研究

目的 研究急性淋巴细胞白血病(ALL)患儿的基因表达谱,探索与儿童ALL危险度相关的新基因.方法 以3例初诊危险度为标危调整后转入高危(B组)和3例初诊和调整后危险度均为标危(A组)的ALL患儿为观察组,3例特发性血小板减少性紫癜患儿为正常对照组,应用Illumina Human-6全基因组表达谱微珠芯片研究B组和A组患儿的差异性基因.通过实时定量RT-PCR,以82例ALL患儿为观察组,21例骨髓象正常患儿为对照组,以ABCC4、BCL11A、TOP2A 3个差异性基因为目的 基因进行验证研究.结果 ①B组与A组患儿有19个基因差异性表达,包括ABCC4、BCL11A 等14个上调基因和TOP2A等5个下调基因.②实时定量RT-PCR检测结果显示ABCC4和BCL11A基因的相对表达量观察组明显高于对照组(P＜0.01);高危组明显高于标危组(P＜0.05);TOP2A基因的相对表达量观察组明显低于对照组(P＜0.01);高危组明显低于标危组(P＜0.05).③与临床指标的相关性研究结果显示,缓解组ABCC4基因的相对表达量明显低于未缓解组(P＜0.05);各组间BCL11A基因的相对表达量差异均无统计学意义(P＞0.05);缓解组TOP2A基因的相对表达量明显高于未缓解组,泼尼松试验敏感组明显高于不敏感组(P＜0.05).结论 儿童ALL的发病及耐药是多基因共同参与的结果,研究ALL患儿的基因表达谱对儿童ALL的耐药机制、预后判断、早期干预和靶向治疗等具有重要意义.

Abstract：

Objective To explore genes associated with risk classification of childhood acute lymphoblastic leukemia(ALL) by gene chip technology. Methods Group A and B were both composed of three newly diagnosed ALL cases with standard risk. After re-evaluation, group B was relegated to high-risk. The control group was composed of three idiopathic thrombocytopenic purpura(ITP) patients. The gene expression profiles of group A and B were studied by Illumina Human-6 Beadchip. Eighty-two ALL patients were selected as the experimental group and 21 with normal bone marrow as control group for real-time quantitative RT-PCR (RQ-PCR). Results ①There were 19 genes expressed differently between group B and A, including 14 upregulated as ABCC4 and BCL11A, 5 down-regulated genes as TOP2A. ②ABCC4 and BCL11 A were validated by RQ-PCR and their expression level was higher in the high risk group than in the standard risk group( P ＜0.05 ). The gene expression level in the group A and B was higher than that in the normal control group( P ＜0.01 ). TOP2A was also validated by RQ-PCR and its expression level in the high risk group was lower than that in the standard risk group(P ＜0.05). The gene expression level in the groups A and B was lower than that in the normal control group and the difference was statistically significance( P ＜ 0. 01 ). ③There was a significant difference in the expression level of ABCC4 between the remission and unremission patients ( P ＜0.05 ). There was no significant difference in the expression level of BCL1 1A between different clinical indicators (P ＞ 0.05). There was significant difference in the expression level of TOP2A between remission and predtnisone good responder groups ( P ＜ 0.05 ). Conclusions Fourteen genes studied were involved in the pathogenesis and drug resistance mechanism in childhood ALL patients. Investigation of gene expression profile will be helpful for predicting drug resistance, prognosis, early intervention and target therapy in childhood ALL.

Preliminary study of gene expression profile associated with risk classification of childhood patients with acute lymphoblastic leukemia

Objective To explore genes associated with risk classification of childhood acute lymphoblastic leukemia(ALL) by gene chip technology. Methods Group A and B were both composed of three newly diagnosed ALL cases with standard risk. After re-evaluation, group B was relegated to high-risk. The control group was composed of three idiopathic thrombocytopenic purpura(ITP) patients. The gene expression profiles of group A and B were studied by Illumina Human-6 Beadchip. Eighty-two ALL patients were selected as the experimental group and 21 with normal bone marrow as control group for real-time quantitative RT-PCR (RQ-PCR). Results ①There were 19 genes expressed differently between group B and A, including 14 upregulated as ABCC4 and BCL11A, 5 down-regulated genes as TOP2A. ②ABCC4 and BCL11 A were validated by RQ-PCR and their expression level was higher in the high risk group than in the standard risk group( P ＜0.05 ). The gene expression level in the group A and B was higher than that in the normal control group( P ＜0.01 ). TOP2A was also validated by RQ-PCR and its expression level in the high risk group was lower than that in the standard risk group(P ＜0.05). The gene expression level in the groups A and B was lower than that in the normal control group and the difference was statistically significance( P ＜ 0. 01 ). ③There was a significant difference in the expression level of ABCC4 between the remission and unremission patients ( P ＜0.05 ). There was no significant difference in the expression level of BCL1 1A between different clinical indicators (P ＞ 0.05). There was significant difference in the expression level of TOP2A between remission and predtnisone good responder groups ( P ＜ 0.05 ). Conclusions Fourteen genes studied were involved in the pathogenesis and drug resistance mechanism in childhood ALL patients. Investigation of gene expression profile will be helpful for predicting drug resistance, prognosis, early intervention and target therapy in childhood ALL.