Nasopharyngeal carcinoma (NPC) is an Epstein-Barr virus (EBV)-associated malignancy with a complex tumor ecosystem. How the interplay between tumor cells, EBV, and the microenvironment contributes to NPC progression and immune evasion remains unclear. Here we performed single-cell RNA sequencing on ~104,000 cells from 19 EBV⁺ NPCs and 7 nonmalignant nasopharyngeal biopsies, simultaneously profiling the transcriptomes of malignant cells, EBV, stromal and immune cells. Overall, we identified global upregulation of interferon responses in the multicellular ecosystem of NPC. Notably, an epithelial–immune dual feature of malignant cells was discovered and associated with poor prognosis. Functional experiments revealed that tumor cells with this dual feature exhibited a higher capacity for tumorigenesis. Further characterization of the cellular components of the tumor microenvironment (TME) and their interactions with tumor cells revealed that the dual feature of tumor cells was positively correlated with the expression of co-inhibitory receptors on CD8⁺ tumor-infiltrating T cells. In addition, tumor cells with the dual feature were found to repress IFN-γ production by T cells, demonstrating their capacity for immune suppression. Our results provide new insights into the multicellular ecosystem of NPC and offer important clinical implications.

鼻咽癌（NPC）是一种与EB病毒（EBV）相关的恶性肿瘤，具有复杂的肿瘤生态系统。肿瘤细胞、EBV 和微环境之间的相互作用如何导致 NPC 进展和免疫逃避仍不清楚。^{在这里，我们对来自 19 个 EBV +}的约 104,000 个细胞进行了单细胞 RNA 测序NPC 和 7 个非恶性鼻咽活检，同时分析恶性细胞、EBV、基质和免疫细胞的转录组。总体而言，我们确定了 NPC 多细胞生态系统中干扰素反应的全球上调。值得注意的是，发现了恶性细胞的上皮-免疫双重特征，并且与预后不良有关。功能实验表明，具有这种双重特征的肿瘤细胞表现出更高的肿瘤发生能力。对肿瘤微环境 (TME) 的细胞成分及其与肿瘤细胞的相互作用的进一步表征表明，肿瘤细胞的双重特征与 CD8 ^{+共抑制受体的表达呈正相关。}肿瘤浸润性 T 细胞。此外，发现具有双重特征的肿瘤细胞可以抑制 T 细胞产生 IFN-γ，证明它们具有免疫抑制能力。我们的研究结果为 NPC 的多细胞生态系统提供了新的见解，并提供了重要的临床意义。