缺氧肿瘤来源的外泌体 miR-21 诱导癌相关成纤维细胞激活促进头颈部鳞状细胞癌转移。

Hypoxic tumor-derived exosomal miR-21 induces cancer-associated fibroblast activation to promote head and neck squamous cell carcinoma metastasis.

机构信息

Department of Maxillofacial and Otorhinolaryngological Oncology, Tianjin Medical University Cancer Institute and Hospital, Key Laboratory of Cancer Prevention and Therapy, Tianjin Cancer Institute, National Clinical Research Center of Cancer, Tianjin 300060, China.

Department of General Surgery, The Second Affiliated Hospital of Anhui Medical University, Hefei 230601, Anhui, China.

出版信息

Cell Signal. 2023 Aug;108:110725. doi: 10.1016/j.cellsig.2023.110725. Epub 2023 May 23.

DOI:10.1016/j.cellsig.2023.110725

PMID:37230199

Abstract

BACKGROUND

Both microRNA-21-5p (miR-21) and the tumor microenvironment, including hypoxia and cancer-associated fibroblasts (CAFs), play a vital role in head and neck squamous cell carcinoma (HNSCC), but whether there is an interaction and the specific regulatory mechanism between them in the process of metastasis is still unclear. In this study, we aimed to elucidate the connection and regulatory mechanism of miR-21, hypoxia, and CAFs in HNSCC metastasis.

METHODS

The underlying mechanisms of hypoxia inducible factor 1 subunit alpha (HIF1α) regulating miR-21 transcription, promoting exosome secretion, CAFs activation, tumor invasion, and lymph node metastasis were determined through quantitative real-time PCR, immunoblotting, transwell, wound healing, immunofluorescence, ChIP, electron microscopy, nanoparticle tracking analysis, dual-luciferase reporter assay, co-culture model and xenografts experiments.

RESULTS

MiR-21 promoted the invasion and metastasis of HNSCC in vitro and in vivo, whereas HIF1α knockdown inhibited these processes. HIF1α upregulated transcription of miR-21 and promoted the release of exosomes from HNSCC cells. Exosomes derived from hypoxic tumor cells were rich in miR-21, which induced NFs activation towards CAFs by targeting YOD1. Knockdown the expression level of miR-21 in CAFs prevented lymph node metastasis in HNSCC.

CONCLUSION

Hypoxic tumor cell-derived exosomal miR-21 might be a therapeutic target to prevent or delay HNSCC invasion and metastasis.

摘要

背景

微小 RNA-21-5p（miR-21）和肿瘤微环境，包括缺氧和癌相关成纤维细胞（CAFs），在头颈部鳞状细胞癌（HNSCC）中起着至关重要的作用，但在转移过程中它们之间是否存在相互作用和具体的调控机制尚不清楚。在这项研究中，我们旨在阐明 miR-21、缺氧和 CAFs 在 HNSCC 转移中的联系和调控机制。

方法

通过定量实时 PCR、免疫印迹、Transwell、划痕、免疫荧光、ChIP、电子显微镜、纳米颗粒跟踪分析、双荧光素酶报告基因检测、共培养模型和异种移植实验，确定缺氧诱导因子 1 亚基α（HIF1α）调节 miR-21 转录、促进外泌体分泌、CAFs 激活、肿瘤侵袭和淋巴结转移的潜在机制。

结果

miR-21 在体外和体内促进了 HNSCC 的侵袭和转移，而 HIF1α 的敲低则抑制了这些过程。HIF1α 上调了 miR-21 的转录，并促进了 HNSCC 细胞中外泌体的释放。来自缺氧肿瘤细胞的外泌体富含 miR-21，通过靶向 YOD1 诱导 NF 激活为 CAFs。在 CAFs 中敲低 miR-21 的表达水平可防止 HNSCC 的淋巴结转移。