Zhang Zhentao, Wang Shuo, Lu Ye, Xia Demeng, Liu Ying
College of Basic Medicine, Naval Medical University, Shanghai 200433, China.
Naval Hospital of Eastern Theater of PLA, Zhoushan, Zhejiang 316000, China.
J Cancer. 2023 Jul 16;14(12):2181-2197. doi: 10.7150/jca.84502. eCollection 2023.
Clear cell renal cell carcinoma (ccRCC) constitutes the commonest kidney malignancy. Immunogenic cell death (ICD) is a type of regulated cell death (RCD), which sufficiently activates adaptive immunity. However, ICD's involvement in cancer development is unclear, as well as the associations of ICD effectors with ccRCC prognosis. RNA-sequencing expression profiles of ccRCC in The Cancer Genome Atlas (TCGA) and normal samples in Gene Expression Omnibus (GEO) were comprehensively investigated. Consensus clustering analysis was employed to determine subgroup members linked to ICD-related genes. Functional enrichment analysis was utilized for the examination of TLR4's biological role, and cellular assays were utilized for further confirmation. We also used Kaplan-Meier (KM) and Cox regression analyses to assess TLR4's prognostic value. Finally, "CIBERSORT" was employed for immune score evaluation. The associations of ICD effectors with ccRCC prognosis were examined based on TCGA, and 12 genes showed upregulation in ccRCC tissue specimens. Meanwhile, ccRCC cases with upregulated ICD-related genes had increased overall survival. Among these ICD-related genes, TLR4 was selected for subsequent analysis. TLR4 was upregulated in ccRCC samples and independently predicted ccRCC. TLR4 also enhanced the proliferative, migratory and invasive abilities in cultured ccRCC cells. Moreover, TLR4 had close relationships with immune checkpoints and infiltrated immune cells. ccRCC cases with elevated TLR4 expression had prolonged overall survival, suggesting a prognostic value for TLR4. Finally, a pan-cancer analysis demonstrated TLR4 had differential expression in various malignancies in comparison with normal tissue samples. This study revealed prognostic values for ICD-associated genes, particularly TLR4, and experimentally validated the inducing effects of TLR4 on ccRCC progression . We also demonstrated the associations of TLR4 with immune cell infiltration, providing a novel strategy for prognostic evaluation and a novel therapeutic target in ccRCC.
透明细胞肾细胞癌（ccRCC）是最常见的肾脏恶性肿瘤。免疫原性细胞死亡（ICD）是一种程序性细胞死亡（RCD），可充分激活适应性免疫。然而，ICD在癌症发展中的作用尚不清楚，ICD效应分子与ccRCC预后的关系也不明确。我们全面研究了癌症基因组图谱（TCGA）中ccRCC的RNA测序表达谱以及基因表达综合数据库（GEO）中的正常样本。采用一致性聚类分析来确定与ICD相关基因相关的亚组成员。利用功能富集分析来研究TLR4的生物学作用，并通过细胞实验进行进一步验证。我们还使用Kaplan-Meier（KM）和Cox回归分析来评估TLR4的预后价值。最后，使用“CIBERSORT”进行免疫评分评估。基于TCGA研究了ICD效应分子与ccRCC预后的关系，12个基因在ccRCC组织标本中上调。同时，ICD相关基因上调的ccRCC病例总生存期延长。在这些与ICD相关的基因中，选择TLR4进行后续分析。TLR4在ccRCC样本中上调，并独立预测ccRCC。TLR4还增强了培养的ccRCC细胞的增殖、迁移和侵袭能力。此外，TLR4与免疫检查点和浸润免疫细胞密切相关。TLR4表达升高的ccRCC病例总生存期延长，提示TLR4具有预后价值。最后，一项泛癌分析表明，与正常组织样本相比，TLR4在各种恶性肿瘤中存在差异表达。本研究揭示了ICD相关基因，特别是TLR4的预后价值，并通过实验验证了TLR4对ccRCC进展的诱导作用。我们还证明了TLR4与免疫细胞浸润的关系，为ccRCC的预后评估提供了新策略和新的治疗靶点。
