Department of Hematology, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Department of Hematology, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Biomed Pharmacother. 2018 Aug;104:181-192. doi: 10.1016/j.biopha.2018.05.047. Epub 2018 May 15.
The long noncoding RNA (lnc) maternally expressed 3 (MEG3) is downregulated in many types of cancers. However, the relationship between lncRNA MEG3, microRNA-21 (miR-21) and chronic myeloid leukemia (CML) blast crisis is unknown. This study examined bone marrow samples from 40 CML patients and 10 healthy donors. Proliferation and apoptosis assays, real-time polymerase chain reaction (PCR), bisulfite sequencing PCR, Western blotting, luciferase assay, RNA pull-down, RNA immunoprecipitation (RIP), co-immunoprecipitation (CoIP) and Chromatin immunoprecipitation (ChIP) were performed. We found that MEG3 and PTEN expression were down-regulated, whereas, MDM2, DNMT1 and miR-21 were up-regulated in the accelerated and blast phases of CML. Treated with 5-azacytidine decreased the level of MDM2, DNMT1 and miR21, but increased the level of MEG3 and PTEN. Overexpression of MEG3 and silencing the expression of miR-21 inhibited proliferation and induced apoptosis. MEG3 overexpression and silencing the expression of miR21 influence the levels of MMP-2, MMP-9, bcl-2 and Bax. MEG3 was able to interact with MDM2 and EZH2. MDM2 could interact with DNMT1 and PTEN. MYC and AKT can interact with EZH2. ChIP-seq showed that the promoter of KLF4 and SFRP2 interacts with DNMT1. In conclusion, lncRNA MEG3 and its target miR21 may serve as novel therapeutic targets for CML blast crisis; and demethylation drugs might also have potential clinical application in treating CML blast crisis.
长链非编码 RNA(lncRNA)母系表达 3(MEG3)在许多类型的癌症中下调。然而,lncRNA MEG3、microRNA-21(miR-21)与慢性髓系白血病(CML)急变期的关系尚不清楚。本研究检测了 40 例 CML 患者和 10 例健康供者的骨髓样本。进行了增殖和凋亡检测、实时聚合酶链反应(PCR)、亚硫酸氢盐测序 PCR、Western blot、荧光素酶检测、RNA 下拉、RNA 免疫沉淀(RIP)、共免疫沉淀(CoIP)和染色质免疫沉淀(ChIP)。结果发现,在 CML 的加速期和急变期,MEG3 和 PTEN 的表达下调,而 MDM2、DNMT1 和 miR-21 的表达上调。用 5-氮杂胞苷处理可降低 MDM2、DNMT1 和 miR21 的水平,但增加 MEG3 和 PTEN 的水平。过表达 MEG3 并沉默 miR-21 的表达可抑制增殖并诱导细胞凋亡。过表达 MEG3 和沉默 miR21 的表达可影响 MMP-2、MMP-9、bcl-2 和 Bax 的水平。MEG3 可与 MDM2 和 EZH2 相互作用。MDM2 可与 DNMT1 和 PTEN 相互作用。MYC 和 AKT 可与 EZH2 相互作用。ChIP-seq 显示 KLF4 和 SFRP2 的启动子与 DNMT1 相互作用。综上所述,lncRNA MEG3 及其靶基因 miR21 可能成为 CML 急变期的新治疗靶点;去甲基化药物也可能具有治疗 CML 急变期的潜在临床应用价值。