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Abstract：
BACKGROUND: Hepatocellular carcinoma (HCC) is a major health concern, necessitating a deeper understanding of its prognosis and underlying mechanisms. This study aimed to investigate the mechanism and prognostic value of CD8+ T Cell exhaustion (CD8+ TEX)-related genes in HCC and construct a survival prognosis prediction model for patients with HCC.
METHODS: CD8+ TEX-related genes associated with HCC prognosis were analysed and identified, and a prognostic prediction model was constructed using the \'least absolute shrinkage and selection operator\' Cox regression model. Immunohistochemistry was used to verify the expression of the model genes in HCC tissues. A nomogram was constructed based on risk scores and clinical features, and its predictive efficacy was verified. The expression of STAM, ANXA5, and MAD2L2 in HCC cell lines was detected by western blotting; subsequently, these genes were knocked down in HCC cell lines by small interfering RNA, and their effects on the proliferation and migration of HCC cell lines were detected by colony formation assay, cck8, wound healing, and transwell assays.
RESULTS: Six genes related to CD8+ TEX were included in the risk-prediction model. The prognosis of patients with HCC in the low-risk group was significantly better than that of those in the high-risk group. Cox regression analysis revealed that the risk score was an independent risk factor for the prognosis of patients with HCC. The differentially expressed genes in patients with high-risk HCC were mainly enriched in the nucleotide-binding oligomerization domain-containing protein-like receptor, hypoxia-inducible factor-1, and tumour programmed cell death protein (PD)-1/PD-L1 immune checkpoint pathways. The CD8+ TEX-related genes STAM, ANXA5, and MAD2L2 were knocked down in HCC cell lines to significantly inhibit cell proliferation and migration. The prediction results of the nomogram based on the risk score showed a good fit and application value.
CONCLUSIONS: The prediction model based on CD8+ TEX-related genes can predict the prognosis of HCC and provide a theoretical basis for the early identification of patients with poor HCC prognosis.
METHODS: CD8+ TEX-related genes associated with HCC prognosis were analysed and identified, and a prognostic prediction model was constructed using the \'least absolute shrinkage and selection operator\' Cox regression model. Immunohistochemistry was used to verify the expression of the model genes in HCC tissues. A nomogram was constructed based on risk scores and clinical features, and its predictive efficacy was verified. The expression of STAM, ANXA5, and MAD2L2 in HCC cell lines was detected by western blotting; subsequently, these genes were knocked down in HCC cell lines by small interfering RNA, and their effects on the proliferation and migration of HCC cell lines were detected by colony formation assay, cck8, wound healing, and transwell assays.
RESULTS: Six genes related to CD8+ TEX were included in the risk-prediction model. The prognosis of patients with HCC in the low-risk group was significantly better than that of those in the high-risk group. Cox regression analysis revealed that the risk score was an independent risk factor for the prognosis of patients with HCC. The differentially expressed genes in patients with high-risk HCC were mainly enriched in the nucleotide-binding oligomerization domain-containing protein-like receptor, hypoxia-inducible factor-1, and tumour programmed cell death protein (PD)-1/PD-L1 immune checkpoint pathways. The CD8+ TEX-related genes STAM, ANXA5, and MAD2L2 were knocked down in HCC cell lines to significantly inhibit cell proliferation and migration. The prediction results of the nomogram based on the risk score showed a good fit and application value.
CONCLUSIONS: The prediction model based on CD8+ TEX-related genes can predict the prognosis of HCC and provide a theoretical basis for the early identification of patients with poor HCC prognosis.
摘要：
背景：肝细胞癌（HCC）是一个主要的健康问题，需要对其预后和潜在机制有更深入的了解。本研究旨在探讨CD8+T细胞耗竭(CD8+TEX)相关基因在HCC中的作用机制及预后价值,构建HCC患者生存预后预测模型。
方法：分析并鉴定与HCC预后相关的CD8+TEX相关基因，并使用“最小绝对收缩和选择算子”Cox回归模型构建了预后预测模型。免疫组织化学用于验证模型基因在HCC组织中的表达。根据风险评分和临床特征构建列线图，并验证了其预测效能。STAM的表达,通过蛋白质印迹法检测HCC细胞系中的ANXA5和MAD2L2；随后，这些基因在肝癌细胞系中被小干扰RNA敲低，集落形成实验检测其对肝癌细胞株增殖和迁移的影响,cck8伤口愈合,和transwell分析。
结果：6个与CD8+TEX相关的基因被纳入风险预测模型。低危组HCC患者的预后明显优于高危组。Cox回归分析显示风险评分是影响HCC患者预后的独立危险因素。高危HCC患者的差异表达基因主要富集在含核苷酸结合寡聚化结构域的蛋白样受体中,缺氧诱导因子-1和肿瘤程序性细胞死亡蛋白（PD）-1/PD-L1免疫检查点途径。CD8+TEX相关基因STAM,在HCC细胞系中敲低ANXA5和MAD2L2以显著抑制细胞增殖和迁移。基于风险评分的列线图预测结果具有较好的拟合效果和应用价值。
结论：基于CD8+TEX相关基因的预测模型可以预测HCC的预后，为早期识别预后不良的HCC患者提供理论依据。
方法：分析并鉴定与HCC预后相关的CD8+TEX相关基因，并使用“最小绝对收缩和选择算子”Cox回归模型构建了预后预测模型。免疫组织化学用于验证模型基因在HCC组织中的表达。根据风险评分和临床特征构建列线图，并验证了其预测效能。STAM的表达,通过蛋白质印迹法检测HCC细胞系中的ANXA5和MAD2L2；随后，这些基因在肝癌细胞系中被小干扰RNA敲低，集落形成实验检测其对肝癌细胞株增殖和迁移的影响,cck8伤口愈合,和transwell分析。
结果：6个与CD8+TEX相关的基因被纳入风险预测模型。低危组HCC患者的预后明显优于高危组。Cox回归分析显示风险评分是影响HCC患者预后的独立危险因素。高危HCC患者的差异表达基因主要富集在含核苷酸结合寡聚化结构域的蛋白样受体中,缺氧诱导因子-1和肿瘤程序性细胞死亡蛋白（PD）-1/PD-L1免疫检查点途径。CD8+TEX相关基因STAM,在HCC细胞系中敲低ANXA5和MAD2L2以显著抑制细胞增殖和迁移。基于风险评分的列线图预测结果具有较好的拟合效果和应用价值。
结论：基于CD8+TEX相关基因的预测模型可以预测HCC的预后，为早期识别预后不良的HCC患者提供理论依据。
