Long non-coding RNA DBCCR1-003 regulate the expression of DBCCR1 via DNMT1 in bladder cancer.
Long non-coding RNA DBCCR1-003 regulate the expression of DBCCR1 via DNMT1 in bladder cancer.
复制标题
长非编码RNA DBCCR1-003通过DNMT1在膀胱癌中调节DBCCR1的表达。
DOI：
10.1186/s12935-016-0356-8
复制
发表时间：
2016
影响因子：
5.8
通讯作者：
Ji W
中科院分区：
文献类型：
--
作者：
Qi D;Li J;Que B;Su J;Li M;Zhang C;Yang M;Zhou G;Ji W
许多长的非编码RNA已被确定为癌症发生过程中的关键调节因子。一个lncRNA，DBCCR1-003，来源于肿瘤抑制基因DBCCR1(缺失于膀胱癌染色体1区)，功能未知。本研究旨在探讨DBCCR1-003在膀胱癌发生发展中的作用及其分子机制。用免疫印迹和实时定量聚合酶链式反应检测DBCCR1-003在组织和细胞中的表达水平。用染色质免疫沉淀法和RNA免疫沉淀法等多种方法证实DBCCR1-003与DNMT1直接结合。构建了高效表达DBCCR1-003的重组载体。采用细胞增殖实验、集落形成实验和流式细胞仪检测DBCCR1-003对细胞增殖、周期和细胞凋亡的调控作用。首先，我们检测了DBCCR1-003、DBCCR1、DNMT1(DNA甲基转移酶1)在DBCCR1启动子中的表达和DNA甲基化。我们发现在BC组织和T24细胞系中，DBCCR1-003低表达，DNMT1高表达，DBCCR1基因启动子高甲基化。进一步的研究表明，DNMT抑制剂5-氮杂-2-脱氧胞苷(DAC)或过表达DBCCR1-003通过逆转启动子超甲基化和DNMT1与DBCCR1启动子的结合而导致T24细胞DBCCR1表达增加。重要的是，RNA免疫沉淀(RIP)表明DBCCR1-003在物理上与DNMT1结合。它们的结合随着DBCCR1启动子甲基化的抑制而增加，表明DBCCR1-003可能与DNMT1结合，并阻止DNMT1介导的DBCCR1甲基化。此外，DBCCR1-003过表达通过诱导G0/G1期停滞和细胞凋亡而显著抑制T24细胞的生长。综上所述，这些发现表明，一种新的肿瘤抑制因子DBCCR1-003通过与DNMT1结合并阻止DNMT1介导的DBCCR1在BC中的甲基化来调节DBCCR1的表达。LncRNA DBCCR1-003有望作为一种新的肿瘤标志物和治疗靶点应用于肿瘤临床。本文的在线版本(doi：10.1186/s12935-0160356-8)包含补充材料，授权用户可以使用。
Many long non coding RNAs have been identified as key modulators in cancer development. A lncRNA, DBCCR1-003, derived from the locus of tumor suppressor gene DBCCR1 (deleted in bladder cancer chromosome region 1), has unknown function. In the present study, we explored function and molecular mechanism of DBCCR1-003 in bladder cancer (BC) development. We evaluated the expression levels of DBCCR1-003 in tissues and cells with western blot and quantitative real-time polymerase chain reaction. Multiple approaches including chromatin immunoprecipitation assay and RNA immunoprecipitation were used to confirm the direct binding of DBCCR1-003 to DNMT1. The recombinant vector overexpressing DBCCR1-003 was constructed. Cell proliferation assay, colony formation assay and flow cytometric analysis were employed to measure the role of DBCCR1-003 in regulation of cell proliferation, cycle and apoptosis. Firstly we detected the expression of DBCCR1-003, DBCCR1, DNMT1 (DNA methyltransferase 1) and DNA methylation in the promoter of DBCCR1. We found low expression of DBCCR1-003, same as DBCCR1, while high expression of DNMT1 and hypermethylation of DBCCR1 gene promoter in BC tissues and T24 cells line. Further studies revealed that treatment of DNMT inhibitor, 5-aza-2-deoxycytidine(DAC), or overexpression of DBCCR1-003 led to increased DBCCR1 expression by reversion of promoter hypermethylation and DNMT1 binding to DBCCR1 promoter in T24 cells. Importantly, RNA immunoprecipitation (RIP) showed that DBCCR1-003 physically associates with DNMT1. The binding of them was increased with the inhibition of DBCCR1 promoter methylation, indicating that DBCCR1-003 may bind to DNMT1 and prevent DNMT1-mediated the methylation of DBCCR1. Furthermore, overexpression of DBCCR1-003 resulted in significant inhibition of T24 cells growth through the inducing G0/G1 arrest and apoptosis. Taken together, these findings demonstrated that a novel tumor suppressor DBCCR1-003 regulates the expression of DBCCR1 via binding to DNMT1 and preventing DNMT1-mediated the methylation of DBCCR1 in BC. LncRNA DBCCR1-003 may serve as a novel biomarker and therapeutic target for BC in future cancer clinic. The online version of this article (doi:10.1186/s12935-016-0356-8) contains supplementary material, which is available to authorized users.
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DOI：
10.1073/pnas.93.18.9821
发表时间：
1996-09-03
影响因子：
11.1
作者：
Herman, JG;Graff, JR;Baylin, SB
通讯作者：
Baylin, SB
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
4.4
作者：
Habuchi, T;Luscombe, N;Knowles, MA
通讯作者：
Knowles, MA
影响因子：
11.2
作者：
Kim, WJ;Kim, EJ;Bae, SC
通讯作者：
Bae, SC
影响因子：
7.7
作者：
Chalei V;Sansom SN;Kong L;Lee S;Montiel JF;Vance KW;Ponting CP
通讯作者：
Ponting CP