TEL-AML1 regulation of survivin and apoptosis via miRNA-494 and miRNA-320a
TEL-AML1 regulation of survivin and apoptosis via miRNA-494 and miRNA-320a
复制标题
DOI：
10.1182/blood-2009-02-206706
复制
发表时间：
2010-12-02
期刊：
影响因子：
20.3
通讯作者：
Wiemels, Joseph L.
中科院分区：
文献类型：
--
作者：
Diakos, Christofer;Zhong, Sheng;Wiemels, Joseph L.
越来越多的证据表明，miRNA和转录因子在正常和恶性造血过程中具有指导意义。我们探讨了儿童白血病中最常见的融合蛋白TEL-AML1 （ETV6-RUNX1）对miRNA表达和白血病表型的影响。通过RNA干扰、miRNA表达阵列和定量聚合酶链反应，我们发现miRNA-494和miRNA-320a在TEL- aml1沉默时上调，而不依赖于TEL的表达。染色质免疫沉淀分析发现miRNA-494是融合蛋白TEL-AML1的直接miRNA靶点。通过生物信息学分析和功能性荧光素酶实验，我们证明了survivin是这2种mirna的靶标。转染miRNA-494和miRNA-320a， Western blot检测survivin表达。这些mirna阻断survivin表达并导致细胞凋亡，其方式与TEL-AML1自身沉默相似；这种沉默也被证明是骰子依赖的。与免疫表型匹配的非TEL-AML1急性淋巴细胞白血病亚型相比，mirna -494和-320a在TEL-AML1+白血病中的表达水平较低，并且在TEL-AML1+白血病中，它们的表达与survivin水平相关。总之，我们的数据表明，TEL-AML1可能至少部分通过抑制miRNA-494和miRNA-320a来发挥其抗凋亡作用，降低它们的表达，从而增强survivin的表达。(血。2010;116 (23):4885 - 4893)
There is increasing evidence that miRNA and transcription factors interact in an instructive fashion in normal and malignant hematopoiesis. We explored the impact of TEL-AML1 (ETV6-RUNX1), the most common fusion protein in childhood leukemia, on miRNA expression and the leukemic phenotype. Using RNA interference, miRNA expression arrays, and quantitative polymerase chain reaction, we identified miRNA-494 and miRNA-320a to be up-regulated upon TEL-AML1 silencing independently of TEL expression. Chromatin immunoprecipitation analysis identified miRNA-494 as a direct miRNA target of the fusion protein TEL-AML1. Using bioinformatic analysis as well as functional luciferase experiments, we demonstrate that survivin is a target of the 2 miRNAs. miRNA-494 and miRNA-320a were introduced to the cells by transfection and survivin expression determined by Western blot analysis. These miRNAs blocked survivin expression and resulted in apoptosis in a similar manner as TEL-AML1 silencing by itself; this silencing was also shown to be Dicer-dependent. miRNAs-494 and -320a are expressed at lower levels in TEL-AML1+ leukemias compared with immunophenotype-matched non TEL-AML1 acute lymphoblastic leukemia subtypes, and within TEL-AML1+ leukemias their expression is correlated to survivin levels. In summary our data suggest that TEL-AML1 might exert its antiapoptotic action at least in part by suppressing miRNA-494 and miRNA-320a, lowering their expression causing enhanced survivin expression. (Blood. 2010;116(23):4885-4893)