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本研究检测RUNX2基因在HOX11+急性T淋巴细胞白血病(T-ALL)细胞株和T-ALL患者骨髓细胞中的甲基化状态和表达水平,探讨其表达水平与启动子区CpG岛甲基化的关系。以3株急性T淋巴细胞白血病细胞株sil-ALL、DND41和RPMI及38例T-ALL患者、29例治疗后完全缓解T-ALL患者骨髓和8例正常人骨髓为样本,采用RT-PCR检测RUNX2 mRNA表达水平,采用重硫酸盐测序法、甲基化DNA免疫沉淀技术和启动子寡核苷酸微阵列芯片分析基因启动子区CpG岛甲基化状态和基因功能。结果表明,高表达HOX11的T-ALL患者样本中会出现RUNX2基因启动子区域的甲基化。RUNX2基因启动子区域在HOX11+T-ALL的甲基化比率为78.9%,明显高于HOX11-ALL(36.8%),两组差异有统计学意义(P<0.01)。RUNX2在HOX11+的细胞株中的表达明显低于HOX11-的细胞株,RUNX2在HOX11+的T-ALL患者中的表达水平(0.581±0.257)明显低于HOX11-的T-ALL患者(0.835±0.317),并且RUNX2 mRNA的相对表达程度与HOX11 mRNA的相对表达程度成负相关(rs=-0.378,P<0.01)。RUNX2基因在T-ALL中的表达水平与其甲基化呈负相关(rs=-0.419,P<0.01)。结论:HOX11能负调控RUNX2的表达,RUNX2基因启动子甲基化是导致其在T-ALL中表达下调或基因沉默的原因,这种HOX11对RUNX2基因的抑制作用有可能就是临床上HOX11高表达T-ALL患者具有更好的无事件生存率和更长的总生存率的原因。RUNX2基因的表达和甲基化水平对判断T-ALL患者的疗效有一定意义,并有望成为一个诊断T-ALL的分子标志。
This study examined the methylation status and expression of RUNX2 gene in bone marrow cells from HOX11 + T-ALL cell lines and T-ALL patients, and explored the relationship between RUNX2 methylation and promoter region CpG island methylation relationship. Thirty-nine acute T-cell leukemia cell lines, sil-ALL, DND41 and RPMI, and 38 T-ALL patients were treated. Twenty-nine patients with complete remission of bone marrow from T-ALL patients and 8 normal human bone marrow samples were used as samples. RT- RUNX2 mRNA expression levels, using methionine DNA immunoprecipitation and promoter oligonucleotide microarray analysis of CpG island methylation status and gene function of gene promoter. The results show that methylation of the promoter region of RUNX2 gene occurs in T-ALL patient samples that are highly expressed HOX11. The methylation ratio of RUNX2 promoter region in HOX11 + T-ALL was 78.9%, which was significantly higher than that in HOX11-ALL (36.8%). There was significant difference between the two groups (P <0.01). The expression of RUNX2 in HOX11 + cell lines was significantly lower than that in HOX11- cells. The expression level of RUNX2 in HOX11 + T-ALL patients (0.581 ± 0.257) was significantly lower than that in HOX11- (0.835 ± 0.317) , And the relative expression of RUNX2 mRNA was negatively correlated with the relative expression of HOX11 mRNA (rs = -0.378, P <0.01). RUNX2 gene expression in T-ALL was negatively correlated with its methylation (rs = -0.419, P <0.01). CONCLUSION: HOX11 can negatively regulate the expression of RUNX2. The promoter methylation of RUNX2 gene causes the down-regulation or gene silencing in T-ALL. The inhibitory effect of HOX11 on RUNX2 gene may be clinically significant T-ALL patients have a better event-free survival and longer overall survival reasons. The expression and methylation of RUNX2 gene have some significance in judging the curative effect of T-ALL patients and are expected to become a molecular marker for diagnosis of T-ALL.