新出现的证据表明,长链非编码RNA(lncRNAs)在各种癌症中起着至关重要的作用。在本研究中,我们的目的是研究上调和生存相关的lncRNA的功能和分子机制,LINC00525,在肺腺癌(LUAD)中。
通过定量逆转录聚合酶链反应(RT-qPCR)和原位杂交(ISH)测定组织中LINC00525的表达水平。LINC00525在LUAD中的功能作用是使用功能增益和损失方法进行研究的,体内和体外。RNA下拉,RNA免疫沉淀(RIP),染色质免疫沉淀(ChIP),三重捕获分析,双荧光素酶测定,基因表达微阵列,和生物信息学分析用于调查潜在的潜在潜在机制。
LINC00525在LUAD细胞和组织中高度表达。生存分析显示LINC00525上调与LUAD患者预后不良相关。在体外敲除LINC00525抑制细胞增殖和细胞周期进程。在异种移植模型中,LINC00525敲除抑制荷瘤小鼠的肿瘤生长和肿瘤发生。机械上,LINC00525通过与p21启动子形成RNA-DNA三链体,将Zeste2聚梳抑制复合物2亚基(EZH2)的增强子引导至p21启动子,从而在表观遗传上抑制p21转录,导致p21启动子的组蛋白3(H3K27me3)上赖氨酸27的三甲基化增加。此外,LINC00525通过增强p21mRNA衰变在转录后抑制p21表达。LINC00525通过竞争性结合RNA结合基序单链相互作用蛋白2(RBMS2)促进p21mRNA衰变。
我们的发现表明,LINC00525通过降低p21mRNA的转录和稳定性来促进LUAD的进展,并与EZH2和RBMS2协同作用,因此表明LINC00525可能是LUAD临床干预的潜在治疗靶标。
Emerging evidence suggests that long noncoding RNAs (lncRNAs) play crucial roles in various cancers. In the present study, we aim to investigate the function and molecular mechanism of an up-regulated and survival-associated lncRNA, LINC00525, in lung adenocarcinoma (LUAD).
The expression level of LINC00525 in tissues was determined by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and in situ hybridization (ISH). The functional role of LINC00525 in LUAD was investigated using gain-and loss-of-function approaches, both in vivo and in vitro. RNA pull-down, RNA immunoprecipitation (RIP), chromatin immunoprecipitation (ChIP), triplex-capture assay, dual-luciferase assay, gene expression microarray, and bioinformatics analysis were used to investigate the potential underlying mechanisms involved.
LINC00525 is highly expressed in LUAD cells and tissues. Survival analysis indicated that upregulation of LINC00525 was associated with poor prognosis in patients with LUAD patients. Knockdown of LINC00525 inhibited cell proliferation and cell cycle progression in vitro. In xenograft models, LINC00525 knockdown suppressed tumor growth and tumorigenesis of tumor-bearing mice. Mechanistically, LINC00525 epigenetically suppressed p21 transcription by guiding Enhancer Of Zeste 2 Polycomb Repressive Complex 2 Subunit (EZH2) to the p21 promoter through an formation of RNA-DNA triplex with the p21 promoter, leading to increased trimethylation of lysine 27 on histone 3 (H3K27me3) of the p21 promoter. In addition, LINC00525 repressed p21 expression post-transcriptionally by enhancing p21 mRNA decay. LINC00525 promoted p21 mRNA decay by competitively binding to RNA Binding Motif Single Stranded Interacting Protein 2 (RBMS2).
Our findings demonstrate that LINC00525 promotes the progression of LUAD by reducing the transcription and stability of p21 mRNA in concert with EZH2 and RBMS2, thus suggesting that LINC00525 may be a potential therapeutic target for clinical intervention in LUAD.