ONECUT1 activates PSAT1 to facilitate PD-L1 expression and mediate immune evasion of lung squamous cell carcinoma

IF 2.5 4区生物学 Q1 ANATOMY & MORPHOLOGY

Tissue & cell Pub Date : 2025-08-15 DOI:10.1016/j.tice.2025.103091

Xiaoyan Huang , Jingwei Yan , Shucong Peng, Yifan Zhou, Shangwei Chen

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引用次数: 0

Abstract

Background

PD-1/PD-L1, a classic immune checkpoint commonly employed in targeted therapy, has proven to yield only limited benefits for patients with lung squamous cell carcinoma (LUSC). Unraveling the intrinsic mechanisms underlying the progression of LUSC serves as the foundation for discovering more effective treatment strategies.

Methods

A study was conducted on the differential expression of PSAT1 and ONECUT1 in LUSC based on data from the TCGA database. Pearson correlation analysis was undertaken to examine the association between PSAT1 expression and CD8⁺ T-cell infiltration or ONECUT1 expression in LUSC. Based on the hTFtarget and JASPAR databases, upstream regulators and potential binding sites for PSAT1 were screened and predicted. An analysis of the prognostic impact of PSAT1 on lung cancer was based on the Kaplan-Meier Plotter website. In qPCR analysis, the mRNA expression of PSAT1 and ONECUT1 was detected. Clinical samples of tumor tissue and adjacent normal tissues were collected, with IHC analysis undertaken on them to determine the PSAT1 protein content, as well as the infiltration level of CD8⁺ T cells in the tumor tissue. Through MTT, cell viability was evaluated. Western blot (WB) was utilized to detect the expression of PD-L1 protein. The ANNEXIN V-FITC experiment was carried out to assess the level of apoptosis in cancer cells. CFSE, ELISA, and Transwell experiments were all employed to assess the activation level and chemotaxis of CD8⁺ T cells. ChIP and dual luciferase assay were carried out to investigate the binding relationship between ONECUT1 and PSAT1.

Results

In tumors and LUSC lines, PSAT1 exhibited significant overexpression, which was linked with adverse prognosis and inversely correlated with CD8⁺ T cell infiltration. In PSAT1-overexpressing tumor tissues, the infiltration level of CD8⁺ T cells was greatly lower than that in tissues with low PSAT1 expression. Knocking down PSAT1 considerably decreased the viability and expression of PD-L1 protein in LUSC cells, as well as upregulated apoptosis levels and the activation and chemotactic capacity of CD8⁺ T cells in co-culture systems. ONECUT1 was a potential upstream regulator of PSAT1, displaying a positive correlation and possessing potential binding sites within the promoter region of PSAT1. Furthermore, ONECUT1 had remarkable overexpression in LUSC tissues and cells. The ChIP and dual luciferase assays confirmed that ONECUT1 effectively bound to PSAT1 and activated PSAT1 expression. Overexpression of ONECUT1 restored the negative regulation on cell viability and positive activation effect on CD8⁺ T cells caused by PSAT1 knockdown.

Conclusion

ONECUT1 transcriptionally activates PSAT1 to reinforce LUSC immune evasion through upregulation of PD-L1 expression.

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ONECUT1激活PSAT1促进PD-L1表达，介导肺鳞状细胞癌的免疫逃避

pd -1/PD-L1是一种经典的免疫检查点，通常用于靶向治疗，已被证明对肺鳞状细胞癌（LUSC）患者的益处有限。揭示LUSC发展的内在机制是发现更有效治疗策略的基础。方法基于TCGA数据库，对LUSC中PSAT1和ONECUT1的差异表达进行研究。采用Pearson相关分析检验PSAT1表达与LUSC中CD8+ t细胞浸润或ONECUT1表达之间的关系。基于hTFtarget和JASPAR数据库，筛选并预测了PSAT1的上游调控因子和潜在结合位点。PSAT1对肺癌预后影响的分析基于Kaplan-Meier Plotter网站。qPCR检测PSAT1和ONECUT1 mRNA表达。收集肿瘤组织及邻近正常组织临床标本，进行免疫组化分析，检测PSAT1蛋白含量及CD8+ T细胞在肿瘤组织中的浸润水平。MTT法测定细胞活力。Western blot （WB）检测PD-L1蛋白的表达。采用ANNEXIN V-FITC实验评估肿瘤细胞凋亡水平。采用CFSE、ELISA和Transwell实验评估CD8+ T细胞的活化水平和趋化性。采用ChIP和双荧光素酶法研究ONECUT1与PSAT1的结合关系。结果PSAT1在肿瘤和LUSC细胞系中显著过表达，与预后不良相关，与CD8+ T细胞浸润呈负相关。在PSAT1过表达的肿瘤组织中，CD8+ T细胞的浸润水平明显低于PSAT1低表达的组织。敲除PSAT1显著降低LUSC细胞中PD-L1蛋白的活力和表达，并上调共培养系统中CD8+ T细胞的凋亡水平和活化和趋化能力。ONECUT1是PSAT1的潜在上游调控因子，与PSAT1的启动子区域呈正相关，并且在PSAT1的启动子区域具有潜在的结合位点。此外，ONECUT1在LUSC组织和细胞中有显著的过表达。ChIP和双荧光素酶实验证实ONECUT1有效结合PSAT1并激活PSAT1表达。过表达ONECUT1恢复了PSAT1敲低对细胞活力的负调控和对CD8+ T细胞的正激活作用。结论onecut1转录激活PSAT1，通过上调PD-L1表达增强LUSC免疫逃避。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Tissue & cell 医学-解剖学与形态学

CiteScore

3.90

自引率

0.00%

发文量

234

期刊介绍： Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed. Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.