A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia.
A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia.
复制标题
一种新型 miR-375-HOXB3-CDCA3/DNMT3B 调节电路有助于急性髓系白血病的白血病发生
DOI：
10.1186/s12885-018-4097-z
复制
发表时间：
2018-02-13
期刊：
影响因子：
3.8
通讯作者：
Gao S
中科院分区：
文献类型：
--
作者：
Bi L;Zhou B;Li H;He L;Wang C;Wang Z;Zhu L;Chen M;Gao S
急性髓性白血病（AML）是一种由复杂的基因突变和表观遗传失调引起的异质性造血恶性肿瘤。MicroRNAs （miRNAs）是一类小的非编码rna，在包括白血病发生在内的所有生物过程中都是重要的基因表达调控因子。最近，miR-375被报道在多种类型的癌症中是一种抑制性miRNA，但其在AML中的潜在抗白血病活性在很大程度上是未知的。方法采用定量逆转录酶PCR （quantitative reverse transcripase PCR, qRT-PCR）检测miR-375和hoxb3在白血病细胞和正常对照中的表达。western blot和荧光素酶检测证实miR-375的靶点。miR-375过表达和HOXB3敲低的表型效应通过生存能力（台盼蓝排斥试验）、菌落形成/复制以及体内肿瘤异种移植试验来评估。结果与正常对照相比，miR-375在白血病细胞系和原代AML细胞中的表达显著降低，这是因为在白血病细胞中发现了前体miR-375 （pre-miR-375）启动子的DNA超甲基化，而在正常对照中没有发现。miR-375的低表达预示着AML患者预后不良。此外，在白血病异种移植小鼠模型中，强迫表达miR-375不仅可以减少白血病细胞的增殖和集落形成，还可以减小异种移植肿瘤的大小，延长存活时间。机制上，miR-375的过表达通过结合hoxb3mrna的3 ' -非翻译区（3 ' -UTR）降低HOXB3的表达并抑制荧光素酶报告基因的活性。过表达HOXB3部分阻断了mir -375诱导的增殖阻滞和集落数量减少，提示HOXB3在mir -375诱导的抗白血病活性中起重要作用。短发夹rna敲低hoxb3可降低细胞分裂周期相关3 （CDCA3）的表达，从而降低细胞增殖。此外，HOXB3诱导DNA甲基转移酶3B （DNMT3B）表达结合在pre-miR-375启动子中，并增强pre-miR-375的DNA超甲基化，导致miR-375的表达降低。总之，我们已经确定了miR-375- hoxb3 - cdca3 /DNMT3B调控回路，它有助于白血病的发生，并提出了一种恢复AML中miR-375表达的治疗策略。
BackgroundAcute myeloid leukemia (AML) is a heterogeneous group of hematopoietic malignancies due to sophisticated genetic mutations and epigenetic dysregulation. MicroRNAs (miRNAs), a class of small non-coding RNAs, are important regulators of gene expression in all biological processes, including leukemogenesis. Recently, miR-375 has been reported to be a suppressive miRNA in multiple types of cancers, but its underlying anti-leukemia activity in AML is largely unknown.MethodsQuantitative reverse transcriptase PCR (qRT-PCR) was used to measure the expression of miR-375 andHOXB3in leukemic cells and normal controls. Targets of miR-375 were confirmed by western blot and luciferase assay. Phenotypic effects of miR-375 overexpression and HOXB3 knockdown were assessed using viability (trypan blue exclusion assay), colony formation/replating, as well as tumor xenograft assays in vivo.ResultsThe expression of miR-375 was substantially decreased in leukemic cell lines and primary AML blasts compared with normal controls, because DNA hypermethylation of precursor-miR-375 (pre-miR-375) promoter was discovered in leukemic cells but not in normal controls. Lower expression of miR-375 predicted poor outcome in AML patients. Furthermore, forced expression of miR-375 not only decreased proliferation and colony formation in leukemic cells but also reduced xenograft tumor size and prolonged the survival time in a leukemia xenograft mouse model. Mechanistically, overexpression of miR-375 reduced HOXB3 expression and repressed the activity of a luciferase reporter through binding 3′-untranslated regions (3’-UTR) ofHOXB3mRNA. Overexpression of HOXB3 partially blocked miR-375-induced arrest of proliferation and reduction of colony number, suggesting that HOXB3 plays an important role in miR-375-induced anti-leukemia activity. Knockdown ofHOXB3by short hairpin RNAs reduced the expression of cell division cycle associated 3 (CDCA3), which decreased cell proliferation. Furthermore, HOXB3 induced DNA methyltransferase 3B (DNMT3B) expression to bind in the pre-miR-375 promoter and enhanced DNA hypermethylation of pre-miR-375, leading to the lower expression of miR-375.ConclusionsCollectively, we have identified a miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry which contributes to leukemogenesis and suggests a therapeutic strategy of restoring miR-375 expression in AML.
登录
查看更多内容
影响因子：
11.4
作者：
Drabkin, H;Parsy, C;Roche, J
通讯作者：
Roche, J
影响因子：
11.4
作者：
Horton, S. J.;Jaques, J.;Schuringa, J. J.
通讯作者：
Schuringa, J. J.
DOI：
10.1016/j.bbrc.2015.10.071
发表时间：
2015-11-27
影响因子：
3.1
作者：
Lindblad, Oscar;Chougule, Rohit A.;Ronnstrand, Lars
通讯作者：
Ronnstrand, Lars
影响因子：
1.6
作者：
Li, Y.;Jiang, Q.;Hu, C.
通讯作者：
Hu, C.
影响因子：
--
作者：
Choi N;Park J;Lee JS;Yoe J;Park GY;Kim E;Jeon H;Cho YM;Roh TY;Lee Y
通讯作者：
Lee Y