Xu Dianqin, Zhou Xinzhu, Min Shaoju, Zhang Yu, Zhu Xiaoyu, Qiao Kun, Xie Luhong, Ren Ji, Liu Yameng, Xiao Ziwen, Tan Yujie
Centre for Clinical Laboratories, The Affiliated Hospital of Guizhou Medical University, Guiyang, 550004, Guizhou, China.
The Affiliated Hospital of Guizhou Medical University, Hypertension Department, Guiyang, 550004, Guizhou, China.
Sci Rep. 2025 Mar 4;15(1):7566. doi: 10.1038/s41598-025-91066-0.
Cervical cancer (CC) is a leading cause of cancer-related mortality among women and is strongly associated with persistent infection by high-risk human papillomavirus (HR-HPV), particularly the HPV16 subtype. Existing detection methods have limitations in meeting clinical requirements. This study aims to identify biomarkers that can aid in the staging and prognosis of cervical cancer. Cervical epithelial exfoliated cell samples were collected from three groups: HPV16-negative normal cervix, HPV16-positive normal cervix, and HPV16-positive cervical cancer. Differential expression proteins (DEPs) were identified using TMT-LC-MS/MS technology, and their associations with tumor-infiltrating lymphocytes (TILs) and immune regulatory molecules were analyzed. Leukocyte-specific protein 1 (LSP1), an intracellular F-actin-binding protein predominantly expressed in macrophages, neutrophils, B cells, and T cells, was identified as a potential biomarker. The expression levels of LSP1 were evaluated and validated using the Human Protein Atlas (HPA) database, immunohistochemistry (IHC), Western blotting (WB), and real-time quantitative PCR (RT-qPCR). Multiplex fluorescence immunohistochemistry (mIHC) was employed to assess the co-localization of LSP1 with CD4 and CD8 T cells. Results indicated that both protein and mRNA levels of LSP1 were significantly elevated in cervical cancer tissues compared to adjacent non-tumor tissues. Notably, LSP1 expression was higher in early-stage cervical cancer (Stage IB) than in advanced-stage disease (Stage IIIC). Furthermore, LSP1 was predominantly localized in CD4 and CD8 T cells with low TIM-3 expression. Analysis of public databases (GEPIA, TIMER2.0, and TISIDB) revealed that higher LSP1 mRNA levels correlated with better patient outcomes. LSP1 expression was positively associated with the abundance of major TILs and immune regulatory molecules, particularly activated B cells, CD8 T cells, and CD4 T cells, while negatively correlated with M2 macrophages and myeloid-derived suppressor cells. These findings indicate that the expression levels of LSP1 in cervical tissues are correlated with cancer staging and patient prognosis, potentially reflecting both tumor immune infiltration and T-cell exhaustion within the tumor microenvironment (TME).
宫颈癌（CC）是女性癌症相关死亡的主要原因，并且与高危型人乳头瘤病毒（HR-HPV）尤其是HPV16亚型的持续感染密切相关。现有的检测方法在满足临床需求方面存在局限性。本研究旨在鉴定有助于宫颈癌分期和预后判断的生物标志物。从三组中收集宫颈上皮脱落细胞样本：HPV16阴性的正常宫颈、HPV16阳性的正常宫颈以及HPV16阳性的宫颈癌。使用TMT-LC-MS/MS技术鉴定差异表达蛋白（DEP），并分析它们与肿瘤浸润淋巴细胞（TIL）和免疫调节分子的关联。白细胞特异性蛋白1（LSP1）是一种主要在巨噬细胞、中性粒细胞、B细胞和T细胞中表达的细胞内F-肌动蛋白结合蛋白，被鉴定为一种潜在的生物标志物。使用人类蛋白质图谱（HPA）数据库、免疫组织化学（IHC）、蛋白质免疫印迹（WB）和实时定量聚合酶链反应（RT-qPCR）对LSP1的表达水平进行评估和验证。采用多重荧光免疫组织化学（mIHC）评估LSP1与CD4和CD8 T细胞的共定位。结果表明，与相邻的非肿瘤组织相比，宫颈癌组织中LSP1的蛋白质和mRNA水平均显著升高。值得注意的是，早期宫颈癌（IB期）中LSP1的表达高于晚期疾病（IIIC期）。此外，LSP1主要定位于TIM-3表达较低的CD4和CD8 T细胞中。对公共数据库（GEPIA、TIMER2.0和TISIDB）的分析表明，较高的LSP1 mRNA水平与更好的患者预后相关。LSP1表达与主要TIL和免疫调节分子的丰度呈正相关，特别是活化的B细胞、CD8 T细胞和CD4 T细胞，而与M2巨噬细胞和髓源性抑制细胞呈负相关。这些发现表明，宫颈组织中LSP1的表达水平与癌症分期和患者预后相关，可能反映了肿瘤微环境（TME）内的肿瘤免疫浸润和T细胞耗竭。
