Department of Stomatology, Liuzhou Worker's Hospital, LiuZhou, Guangxi Zhuang Autonomous Region, China.
Department of Stomatology, Fourth Affiliated Hospital of Guangxi Medical University, LiuZhou, Guangxi Zhuang Autonomous Region, China.
Sci Rep. 2024 Nov 27;14(1):29458. doi: 10.1038/s41598-024-79417-9.
Recent studies have identified SLC2A3 as being abnormally upregulated in multiple tumor types, correlating with poor survival and disrupted microenvironments. However, its prognostic significance in head and neck squamous cell carcinoma (HNSC) remains underexplored. In this study, SLC2A3 was screened as a potential risk gene influencing both immune and tumor components within the tumor microenvironment (TME) of 504 HNSC patients from the TCGA database. Immune infiltration analyses and clinical significance on SLC2A3 were conducted using ESTIMATE, CIBERSORT, ssGSEA, TIMER and clinical prognosis parameters. Additionally, the single-cell dataset is used to analyze the expression of SLC2A3 in various subpopulations. The magnetic activated cell sorting (MACS) is used to isolate CD8 T cells from PBMCs or tumor tissues. Flow cytometry is used to identify purified and activated CD8 T cells. GSEA and WB were used to investigate the molecular mechanism of SLC2A3 in CD8 T cells. The co-culture system of CD8 T cells and TU686 was used to investigate the effects of SLC2A3 on immune cells and tumor development. In this study, SLC2A3 was identified as a potential risk gene affecting both immune cells and tumor components within the TME of 504 HNSC patients derived from the TCGA database. We conducted immune infiltration analyses and assessed the clinical significance of SLC2A3 using various bioinformatics tools, including ESTIMATE, CIBERSORT, ssGSEA, and TIMER, along with clinical prognosis parameters. The single-cell RNA sequencing dataset was utilized to examine SLC2A3 expression across different cellular subpopulations. Magnetic activated cell sorting (MACS) was employed to isolate CD8 T cells from peripheral blood mononuclear cells (PBMCs) or tumor tissues. Flow cytometry was implemented to confirm the purity and activation state of the isolated CD8 T cells. GSEA and Western blot were applied to explore the molecular mechanisms underlying SLC2A3's role in CD8 T cells. Lastly, a co-culture system involving CD8 T cells and TU686 tumor cells was established to study the impact of SLC2A3 on immune cell function and tumor progression. SLC2A3 emerges as an actively variable gene within the immune and stromal components of the TME, linked to aggravated immune infiltration and poor clinical outcomes. The upregulated expression of SLC2A3 is predominantly enriched in immune-related biological processes and linked to the suppression of CD8 T cells, which are crucial for the survival of HNSC patients. Furthermore, SLC2A3 exhibits specific overexpression in CD8 T cells and may potentially trigger ferroptosis. Knockdown of SLC2A3 led to a significant increase in the proliferation of CD8 T cells compared to those without knockdown. In co-culture systems, CD8 T cells with SLC2A3 knockdown demonstrated an enhanced ability to eliminate tumor cells compared to those without the knockdown. SLC2A3 is associated with changes in the TME and prognostic indicators. Moreover, high SLC2A3 expression in CD8 T cells may drive cell death through ferroptosis, fostering tumor progression.
最近的研究表明，SLC2A3 在多种肿瘤类型中异常上调，与不良预后和微环境破坏有关。然而，其在头颈部鳞状细胞癌（HNSC）中的预后意义仍未得到充分探索。在这项研究中，我们从 TCGA 数据库中筛选了 504 例 HNSC 患者的肿瘤微环境（TME）中的潜在风险基因 SLC2A3，该基因可能同时影响免疫和肿瘤成分。使用 ESTIMATE、CIBERSORT、ssGSEA、TIMER 和临床预后参数进行 SLC2A3 的免疫浸润分析和临床意义分析。此外，还使用单细胞数据集分析 SLC2A3 在各种亚群中的表达。使用磁激活细胞分选（MACS）从外周血单核细胞（PBMCs）或肿瘤组织中分离 CD8+T 细胞。使用流式细胞术鉴定纯化和激活的 CD8+T 细胞。使用 GSEA 和 WB 研究 SLC2A3 在 CD8+T 细胞中的分子机制。使用 CD8+T 细胞和 TU686 的共培养系统研究 SLC2A3 对免疫细胞和肿瘤发展的影响。
在这项研究中，我们从 TCGA 数据库中筛选了 504 例 HNSC 患者的肿瘤微环境（TME）中的潜在风险基因 SLC2A3，该基因可能同时影响免疫和肿瘤成分。我们使用各种生物信息学工具，包括 ESTIMATE、CIBERSORT、ssGSEA 和 TIMER 以及临床预后参数进行 SLC2A3 的免疫浸润分析和临床意义评估。使用单细胞 RNA 测序数据集分析 SLC2A3 在不同细胞亚群中的表达。使用磁激活细胞分选（MACS）从外周血单核细胞（PBMCs）或肿瘤组织中分离 CD8+T 细胞。使用流式细胞术鉴定分离的 CD8+T 细胞的纯度和激活状态。使用 GSEA 和 WB 研究 SLC2A3 在 CD8+T 细胞中的分子机制。最后，建立了 CD8+T 细胞和 TU686 肿瘤细胞的共培养系统，研究 SLC2A3 对免疫细胞功能和肿瘤进展的影响。
SLC2A3 是 TME 中免疫和基质成分中一个活跃变化的基因，与加重的免疫浸润和不良的临床结局有关。SLC2A3 的上调表达主要富集在与免疫相关的生物学过程中，并与 CD8+T 细胞的抑制有关，而 CD8+T 细胞的抑制与 HNSC 患者的生存有关。此外，SLC2A3 在 CD8+T 细胞中特异性过表达，并可能潜在地触发铁死亡。与未敲低的细胞相比，敲低 SLC2A3 导致 CD8+T 细胞的增殖显著增加。在共培养系统中，与未敲低的细胞相比，敲低 SLC2A3 的 CD8+T 细胞具有更强的消除肿瘤细胞的能力。SLC2A3 与 TME 和预后指标的变化有关。此外，SLC2A3 在 CD8+T 细胞中的高表达可能通过铁死亡导致细胞死亡，促进肿瘤进展。
