Wu Kainan, Chang Fen, Li Wenming, Wei Dongmin, Cao Shengda, Xie Yulin, Li Ce, Lei Dapeng
Department of Otorhinolaryngology, Qilu Hospital of Shandong University, NHC Key Laboratory of Otorhinolaryngology (Shandong University), Jinan, Shandong, China.
Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.
Front Oncol. 2023 Apr 25;13:1117622. doi: 10.3389/fonc.2023.1117622. eCollection 2023.
Hypopharyngeal squamous cell cancer (HSCC) is one of the most malignant tumors of the head and neck. It is not easy to detect in the early stage due to its hidden location; thus, lymph node metastasis is highly likely at diagnosis, leading to a poor prognosis. It is believed that epigenetic modification is related to cancer invasion and metastasis. However, the role of m6A-related lncRNA in the tumor microenvironment (TME) of HSCC remains unclear.
The whole transcriptome and methylation sequencing of 5 pairs of HSCC tissues and adjacent tissues were performed to identify the methylation and transcriptome profiles of lncRNAs. The biological significance of lncRNAs differentially expressing the m6A peak was analyzed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes. By constructing an m6A lncRNA-microRNA network, the mechanism of m6A lncRNAs in HSCC was analyzed. The relative expression levels of selected lncRNAs were examined by quantitative polymerase chain reaction. The CIBERSORT algorithm was used to evaluate the relative proportion of immune cell infiltration in HSCC and paracancerous tissues.
Based on an in-depth analysis of the sequencing results, 14413 differentially expressed lncRNAs were revealed, including 7329 up-regulated and 7084 down-regulated lncRNAs. Additionally, 4542 up-methylated and 2253 down-methylated lncRNAs were detected. We demonstrated methylation patterns and gene expression profiles of lncRNAs of HSCC transcriptome. In the intersection analysis of lncRNAs and methylated lncRNAs, 51 lncRNAs with up-regulated transcriptome and methylation and 40 lncRNAs with down-regulated transcriptome and methylation were screened, and significantly differentiated lncRNAs were further studied. In the immune cell infiltration analysis, B cell memory was significantly elevated in cancer tissue, while γδT cell amount was significantly decreased.
m6A modification of lncRNAs might be involved in HSCC pathogenesis. Infiltration of immune cells in HSCC might provide a new direction for its treatment. This study provides new insights for exploring the possible HSCC pathogenesis and searching for new potential therapeutic targets.
下咽鳞状细胞癌（HSCC）是头颈部最恶性的肿瘤之一。由于其位置隐匿，早期不易被发现；因此，在诊断时淋巴结转移的可能性很高，导致预后较差。据信表观遗传修饰与癌症侵袭和转移有关。然而，m6A相关lncRNA在HSCC肿瘤微环境（TME）中的作用仍不清楚。
对5对HSCC组织和癌旁组织进行全转录组和甲基化测序，以鉴定lncRNAs的甲基化和转录组图谱。通过基因本体论和京都基因与基因组百科全书分析差异表达m6A峰的lncRNAs的生物学意义。通过构建m6A lncRNA-微小RNA网络，分析m6A lncRNAs在HSCC中的作用机制。通过定量聚合酶链反应检测所选lncRNAs的相对表达水平。使用CIBERSORT算法评估HSCC和癌旁组织中免疫细胞浸润的相对比例。
通过对测序结果的深入分析，共鉴定出14413个差异表达的lncRNAs，其中7329个上调，7084个下调。此外，检测到4542个甲基化上调和2253个甲基化下调的lncRNAs。我们展示了HSCC转录组lncRNAs的甲基化模式和基因表达谱。在lncRNAs与甲基化lncRNAs的交集分析中，筛选出51个转录组和甲基化上调的lncRNAs和40个转录组和甲基化下调的lncRNAs，并对显著差异的lncRNAs进行进一步研究。在免疫细胞浸润分析中，癌组织中B细胞记忆显著升高，而γδT细胞数量显著减少。
lncRNAs的m6A修饰可能参与HSCC的发病机制。HSCC中免疫细胞的浸润可能为其治疗提供新的方向。本研究为探索HSCC可能的发病机制和寻找新的潜在治疗靶点提供了新的见解。
