LINC01419 通过招募 EZH2 并调节 FBP1 表达来调控肺腺癌中的细胞干性。
The role of LINC01419 in regulating the cell stemness in lung adenocarcinoma through recruiting EZH2 and regulating FBP1 expression.
机构信息
Department of Thoracic Surgery, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Qingchun Road East, Hangzhou, 310016, Zhejiang Province, China.
出版信息
Biol Direct. 2022 Sep 1;17(1):23. doi: 10.1186/s13062-022-00336-8.
BACKGROUND
Recent years have witnessed a growing academic interest in the effects of lncRNAs on tumors. LINC01419 is found to facilitate proliferation and metastasis of lung adenocarcinoma (LUAD) cells, but there is a great deal of uncertainty about how LINC01419 works on LUAD cell stemness. For this reason, the focus of this research is centered on the regulatory impact of LINC01419 on LUAD cell stemness.
METHODS
For the detection of the expression level of LINC01419 in LUAD, qRT-PCR was performed. And how oe-LINC01419 and sh-LINC01419 affected LUAD cell proliferation as well as stem cell sphere-formation were examined by CCK-8 and cell sphere-forming assays. In addition, whether LINC01419 could recruit EZH2 and regulate FBP1 expression were determined by bioinformatics analysis, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (ChIP). Western blot was utilized to detect the protein expression levels of FBP1, CD44, CD133, and ALDH-1 as well.
RESULTS
On the basis of the findings from those assays, an up-regulation of LINC01419 level was demonstrated in LUAD cell lines, and a remarkable upregulation of it in CD44 + LUAD cells. In LUAD cells, proliferation and stem cell sphere-formation that were attenuated by LINC01419 knockdown were discovered to be facilitated by LINC01419 overexpression. And a binding relationship between LINC01419 and EZH2 was determined by RIP assay. Besides, EZH2 was capable of binding to FBP1 promoter region, as found by ChIP-PCR assay. Finally, it was demonstrated by in vitro experiments that LINC01419 could inhibit FBP1 expression by recruiting EZH2, resulting in promotion of LUAD cell proliferation and stemness.
SIGNIFICANCE
To summarize, our findings demonstrate a cancer-promoting role of LINC01419 in LUAD. LINC01419, by recruiting EZH2 and regulating FBP1 expression, contributes to LUAD cell stemness. According to these findings, the potential of LINC01419 to be the target for LUAD treatment is hence determined, which also adds more possibility to the enrichment of therapeutic strategies for lung cancer stem cells.
背景
近年来，人们对长链非编码 RNA（lncRNA）对肿瘤影响的学术兴趣日益浓厚。研究发现 LINC01419 可促进肺腺癌（LUAD）细胞的增殖和转移，但 LINC01419 如何影响 LUAD 细胞干性仍存在很大的不确定性。因此，本研究的重点是集中研究 LINC01419 对 LUAD 细胞干性的调控作用。
方法
通过 qRT-PCR 检测 LUAD 中 LINC01419 的表达水平。通过 CCK-8 检测 oe-LINC01419 和 sh-LINC01419 对 LUAD 细胞增殖的影响，通过细胞球形成实验检测干细胞球形成。此外，通过生物信息学分析、RNA 免疫沉淀（RIP）和染色质免疫沉淀（ChIP）实验来确定 LINC01419 是否可以募集 EZH2 并调节 FBP1 的表达。Western blot 检测 FBP1、CD44、CD133 和 ALDH-1 的蛋白表达水平。
结果
基于这些实验结果，发现在 LUAD 细胞系中 LINC01419 水平上调，在 CD44+LUAD 细胞中上调更为显著。在 LUAD 细胞中，LINC01419 敲低减弱的增殖和干细胞球形成能力，通过 LINC01419 过表达得到促进。通过 RIP 实验确定了 LINC01419 与 EZH2 之间的结合关系。此外，通过 ChIP-PCR 实验发现 EZH2 能够结合到 FBP1 启动子区域。最后，通过体外实验证明 LINC01419 通过募集 EZH2 抑制 FBP1 的表达，从而促进 LUAD 细胞的增殖和干性。
意义
总之，我们的研究结果表明，LINC01419 在 LUAD 中发挥促癌作用。LINC01419 通过募集 EZH2 并调节 FBP1 的表达，促进 LUAD 细胞干性。根据这些发现，LINC01419 有可能成为 LUAD 治疗的靶点，也为肺癌干细胞治疗策略的丰富提供了更多的可能性。
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