Department of Otolaryngology-Head and Neck Surgery, Gunma University Graduate School of Medicine, Japan.
Department of Otolaryngology-Head and Neck Surgery, Gunma University Graduate School of Medicine, Japan.
Oral Oncol. 2023 Feb;137:106296. doi: 10.1016/j.oraloncology.2022.106296. Epub 2022 Dec 24.
Human papillomavirus (HPV)-positive head and neck squamous cell carcinoma (HNSCC) differs in terms of cellular and molecular biological characteristics from HPV-negative HNSCC. However, differences in circulating tumor cells (CTCs) between HPV-positive and -negative HNSCC remain unclear.
We first analyzed eight epithelial-mesenchymal transition (EMT)-related genes (VIM, CDH1, CDH2, SNAI1, SNAI2, TWIST1, ZEB1, and ZEB2) using The Cancer Genome Atlas (TCGA) database. Next, we isolated CTCs from patients with HNSCC using CD45-negative selection and expression analysis of epithelial-related genes (EPCAM, EGFR, and MET) by RT-qPCR. CTC-positive samples were further analyzed for EMT-related genes. In addition, we investigated the proportion of circulating T cell subsets and CD38+ T cells using flow cytometry and their involvement in CTCs.
Compared with HPV-negative HNSCC, expression of CDH1, SNAI1, SNAI2, TWIST1, and ZEB1 was downregulated in HPV-positive HNSCC, as determined by TCGA analysis. CTCs were detected in 19 (52.8 %) of 36 HPV-positive and 26 (68.4 %) of 38 HPV-negative patients with HNSCC. EPCAM-positive and MET-positive CTCs were significantly more frequent in patients with HPV-negative HNSCC. HPV-positive patients with HNSCC exhibited significantly high SNAI1 and ZEB2 expression in CTCs. Interestingly, differences in SNAI1 expression levels differed markedly between CTCs and TCGA based on HPV status. Moreover, HPV-positive patients with HNSCC exhibiting SNAI1-high CTCs showed a superior prognosis and a lower proportion of CD38+ T cells than those with SNAI1-low CTCs.
Our results provide novel insights into the EMT-MET spectrum of CTCs and may contribute to the development of prognostic biomarkers for HPV-positive HNSCC.
人乳头瘤病毒（HPV）阳性头颈部鳞状细胞癌（HNSCC）在细胞和分子生物学特征上与 HPV 阴性 HNSCC 不同。然而，HPV 阳性和阴性 HNSCC 之间循环肿瘤细胞（CTC）的差异尚不清楚。
我们首先使用癌症基因组图谱（TCGA）数据库分析了 8 个上皮间质转化（EMT）相关基因（VIM、CDH1、CDH2、SNAI1、SNAI2、TWIST1、ZEB1 和 ZEB2）。接下来，我们使用 CD45 阴性选择法从 HNSCC 患者中分离 CTCs，并通过 RT-qPCR 分析上皮相关基因（EPCAM、EGFR 和 MET）的表达。CTC 阳性样本进一步进行 EMT 相关基因分析。此外，我们通过流式细胞术研究了循环 T 细胞亚群和 CD38+T 细胞的比例及其与 CTC 的关系。
通过 TCGA 分析，与 HPV 阴性 HNSCC 相比，HPV 阳性 HNSCC 中 CDH1、SNAI1、SNAI2、TWIST1 和 ZEB1 的表达下调。在 36 例 HPV 阳性和 38 例 HPV 阴性 HNSCC 患者中，检测到 19 例（52.8%）和 26 例（68.4%）患者存在 CTCs。EPCAM 阳性和 MET 阳性 CTCs 在 HPV 阴性 HNSCC 患者中更为常见。HPV 阳性 HNSCC 患者的 CTC 中 SNAI1 和 ZEB2 表达明显较高。有趣的是，基于 HPV 状态，CTC 与 TCGA 之间 SNAI1 表达水平的差异非常显著。此外，HPV 阳性 HNSCC 患者的 SNAI1 高 CTCs 具有更好的预后，且 CD38+T 细胞比例较低。
我们的研究结果为 CTC 的 EMT-MET 谱提供了新的见解，可能有助于开发 HPV 阳性 HNSCC 的预后生物标志物。
