Chen Hai-Bin, Gong Xiao-Yang, Shen Wen-Hao, Zhu Zi-Hang, Chen Xi
Department of Otorhinolaryngology, The First Affiliated Hospital, Nanjing Medical University, Nanjing, 210029, China.
Heliyon. 2024 Aug 15;10(16):e36358. doi: 10.1016/j.heliyon.2024.e36358. eCollection 2024 Aug 30.
Long non-coding RNAs (lncRNAs) are closely associated with the pathogenesis of laryngeal squamous cell carcinoma (LSCC). This study aimed to investigate the roles of AC068768.1 in LSCC.
Exosomes were extracted by ultracentrifugation and identified by transmission electron microscopy (TEM) assay. The expression levels of mRNA and miRNA were determined by real-time quantitative polymerase chain reaction (RT-qPCR). Cellular functions were assesses through immunofluorescence, flow cytometry, colony formation, wound healing and transwell assays. Chromatin immunoprecipitation (ChIP) and luciferase assays were conducted to verify the binding of AC068768.1 by signal transducer and activator of transcription 3 (STAT3). Xenograft assays were performed to confirm the roles of AC068768.1 in LSCC, and hematoxylin-eosin (HE) staining was applied for histological analysis.
LSCC cell-derived exosomes induced M2-like tumor-associated macrophages (TAM2) polarization, which promoted the proliferation, migration, and invasion of LSCCs. Knockdown of exosomal AC068768.1 inhibited M2 polarization and suppressed LSCC aggressiveness both in vitro and in vivo. Moreover, AC068768.1 sponged miR-139-5p, inducing the upregulation of neurogenic locus notch homolog protein 1 (NOTCH1). LSCCs adapted to TAM2 polarization in the tumor microenvironment via AC068768.1-mediated activation of the NOTCH1 pathway. Additionally, NOTCH1 activated STAT3.
The AC068768.1/miR-139-5p/NOTCH1/STAT3 axis promotes the metastasis of LSCC. This finding may provide a novel target for LSCC therapy.
长链非编码RNA（lncRNAs）与喉鳞状细胞癌（LSCC）的发病机制密切相关。本研究旨在探讨AC068768.1在LSCC中的作用。
通过超速离心提取外泌体，并通过透射电子显微镜（TEM）检测进行鉴定。通过实时定量聚合酶链反应（RT-qPCR）测定mRNA和miRNA的表达水平。通过免疫荧光、流式细胞术、集落形成、伤口愈合和Transwell检测评估细胞功能。进行染色质免疫沉淀（ChIP）和荧光素酶检测以验证信号转导和转录激活因子3（STAT3）与AC068768.1的结合。进行异种移植检测以证实AC068768.1在LSCC中的作用，并应用苏木精-伊红（HE）染色进行组织学分析。
LSCC细胞来源的外泌体诱导M2样肿瘤相关巨噬细胞（TAM2）极化，促进LSCC的增殖、迁移和侵袭。外泌体AC068768.1的敲低抑制了M2极化，并在体外和体内抑制了LSCC的侵袭性。此外，AC068768.1吸附miR-139-5p，导致神经源性位点Notch同源蛋白1（NOTCH1）上调。LSCC通过AC068768.1介导的NOTCH1途径激活在肿瘤微环境中适应TAM2极化。此外，NOTCH1激活STAT3。
AC068768.1/miR-139-5p/NOTCH1/STAT3轴促进LSCC的转移。这一发现可能为LSCC治疗提供新的靶点。
