臭椿酮通过胶质母细胞瘤中的HIF1-α/LINC01956/FUS/β-连环蛋白信号通路干扰癌细胞与肿瘤相关巨噬细胞之间的串扰。

Ailanthone disturbs cross-talk between cancer cells and tumor-associated macrophages via HIF1-α/LINC01956/FUS/β-catenin signaling pathway in glioblastoma.

作者信息

Deng Xubin, Zhang Qianbing, Jin Fa, Lu Fengfei, Duan Guosheng, Han Luwei, Zhu Meiling, Yang Zhengyan, Zhang Gong

机构信息

Guangzhou Institute of Cancer Research, the Affiliated Cancer Hospital, Guangzhou Medical University, Guangzhou, China.

Cancer Research Institute, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.

出版信息

Cancer Cell Int. 2024 Dec 5;24(1):397. doi: 10.1186/s12935-024-03594-w.

DOI:10.1186/s12935-024-03594-w

PMID:39639311

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11619249/

Abstract

BACKGROUND

An increasing number of studies have focused on ailanthone (aila) due to its antitumor activity. However, the role of ailanthone in glioblastoma(GBM) has not been investigated before. This study aims to explore the biological function and the underlying mechanism of ailanthone in GBM.

METHODS

The microarray analysis was used to screen out down-stream long non-coding RNAs (lncRNAs) targeted by ailanthone. Real-time PCR(RT-PCR) assay was used to examine LINC01956 expression levels. Colony-formation, Methylthiazolyldiphenyl-tetrazolium bromide(MTT), cell-cycle, organoids culture and in-vivo tumorigenesis assays were used to examine cell growth in vitro and in vivo. Boyden assay was used to examine cell invasion ability in vitro. RNA immunoprecipitation and RNA-protein pull-down assays were used to examine the interaction between LINC01956 and FUS protein. Chromatin Immunoprecipitation(ChIP) assay was used to examine HIF1-α-binding sites in the LINC01956 promoter.

RESULTS

Ailanthone decreased GBM cell growth in vitro and in vivo via inducing ferroptosis. Ailanthone treatment exhibited blood‒brain barrier(BBB) permeability and specifically targeted the tumor area. LINC01956 was identified as a down-stream target of Ailanthone. LINC01956 exerted as an onco-lncRNA in GBM. M2 polarization of macrophages induced by exosomes derived from glioma cells overexpressing LINC01956 accelerated GBM progression. Mechanistically, we found that LINC01956 bound to FUS and reduced its ubiquitination. LINC01956 evoked nuclear translocation of phosphorylated (p)-β-catenin by recruiting FUS. Furthermore, under hypoxic conditions, LINC01956 was regulated by HIF-1α. Ailanthone decreased the expression of LINC01956 via suppressing HIF-1α.

CONCLUSION

Taken together, our data revealed for the first time that ailanthone regulated HIF-1α/LINC01956/FUS/β-catenin signaling pathway and thereby inhibited GBM progression.

摘要

背景

由于其抗肿瘤活性，越来越多的研究聚焦于臭椿酮（aila）。然而，臭椿酮在胶质母细胞瘤（GBM）中的作用此前尚未被研究。本研究旨在探索臭椿酮在GBM中的生物学功能及潜在机制。

方法

采用微阵列分析筛选臭椿酮靶向的下游长链非编码RNA（lncRNA）。运用实时定量聚合酶链反应（RT-PCR）检测LINC01956的表达水平。采用集落形成、甲基噻唑基二苯基四氮唑溴盐（MTT）、细胞周期、类器官培养及体内肿瘤发生实验检测体外和体内的细胞生长情况。采用博伊登实验检测体外细胞侵袭能力。运用RNA免疫沉淀和RNA-蛋白质下拉实验检测LINC01956与FUS蛋白之间的相互作用。采用染色质免疫沉淀（ChIP）实验检测LINC01956启动子中的缺氧诱导因子1α（HIF1-α）结合位点。

结果

臭椿酮通过诱导铁死亡降低GBM细胞的体外和体内生长。臭椿酮治疗具有血脑屏障（BBB）通透性，并特异性靶向肿瘤区域。LINC01956被鉴定为臭椿酮的下游靶点。LINC01956在GBM中作为一种致癌lncRNA发挥作用。过表达LINC01956的胶质瘤细胞来源的外泌体诱导的巨噬细胞M2极化加速了GBM的进展。机制上，我们发现LINC01956与FUS结合并降低其泛素化。LINC01956通过招募FUS引发磷酸化（p）-β-连环蛋白的核转位。此外，在缺氧条件下，LINC01956受HIF-1α调控。臭椿酮通过抑制HIF-1α降低LINC01956的表达。