Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, NHC Key Laboratory of Glycoconjugates Research (Fudan University), Shanghai, PR China.
Zhejiang Provincial People's Hospital, Hangzhou, PR China.
Life Sci. 2021 Sep 1;280:119748. doi: 10.1016/j.lfs.2021.119748. Epub 2021 Jun 24.
BACKGROUND & AIMS: Hepatocellular carcinoma (HCC) is the most common primary malignancy of the liver. Long non-coding RNAs as master gene regulators play important roles in tumorigenesis and progression. However, the significance of lncRNAs and their regulatory mechanisms in HCC are largely unknown. Our study was to define the role of lncAY (long noncoding RNA AY927503) in HCC.
Methylated RNA immunoprecipitation qPCR combined with bioinformatics were used to identify the m6A modification of lncAY. qRT-PCR, western blotting and immunofluorescence were used to identify the expression of the lncAY/YTHDF2/BMI1/Wnt axis in HCC tissues and cell lines. Gain- and loss-of functions of lncAY and BMI1 were implemented to confirm their roles in the behaviors of HCC cells.
Our findings suggested that m6A-modified lncAY expression relied on m6A "reader" protein YTHDF2. LncAY upregulated BMI1 expression in HCC cells and a notably positive relevance is evident between lncAY and BMI1 expression in TCGA HCC datasets. BMI1 was upregulated in HCC tissues and patients with higher BMI1 expression had a poor clinical prognosis. Besides, GSEA analysis showed remarkable enrichment of high BMI1 expression in gene sets associated with Wnt/β-catenin signaling. Rescue results revealed that BMI1 reversed the suppressive effects of lncAY depletion in HCC cells.
Our work suggested that lncAY might elevate BMI1 expression and further activate the Wnt/β-catenin signaling. BMI1 reverses the suppressive effects of lncAY depletion in HCC cells. Collectively, our work uncovers a novel undefined regulatory signaling pathway, namely lncAY/BMI1/Wnt/β-catenin axis, involved in liver cancer progression.
肝细胞癌（HCC）是肝脏最常见的原发性恶性肿瘤。长链非编码 RNA 作为主基因调控因子，在肿瘤发生和发展中发挥重要作用。然而，lncRNAs 的意义及其在 HCC 中的调控机制在很大程度上尚不清楚。本研究旨在定义 lncAY（AY927503 长链非编码 RNA）在 HCC 中的作用。
采用 m6A 修饰 RNA 免疫沉淀 qPCR 结合生物信息学方法鉴定 lncAY 的 m6A 修饰。qRT-PCR、western blot 和免疫荧光法用于鉴定 HCC 组织和细胞系中 lncAY/YTHDF2/BMI1/Wnt 轴的表达。实施 lncAY 和 BMI1 的增益和缺失功能，以确认它们在 HCC 细胞行为中的作用。
我们的研究结果表明，m6A 修饰的 lncAY 表达依赖于 m6A“阅读器”蛋白 YTHDF2。lncAY 在 HCC 细胞中上调 BMI1 的表达，在 TCGA HCC 数据集，lncAY 和 BMI1 的表达之间存在显著的正相关性。BMI1 在 HCC 组织中上调，且具有较高 BMI1 表达的患者具有较差的临床预后。此外，GSEA 分析显示，高 BMI1 表达在与 Wnt/β-catenin 信号相关的基因集中有显著富集。挽救结果表明，BMI1 逆转了 lncAY 耗竭对 HCC 细胞的抑制作用。
我们的研究结果表明，lncAY 可能通过上调 BMI1 表达，进而激活 Wnt/β-catenin 信号通路。BMI1 逆转了 lncAY 耗竭对 HCC 细胞的抑制作用。总之，我们的工作揭示了一个新的未知的调控信号通路，即 lncAY/BMI1/Wnt/β-catenin 轴，参与肝癌的进展。
Zotero 插件