Chronic inflammation is involved in the initiation and progression of various cancers, including liver cancer. The current study focuses on the characterization of the peripheral immune response in hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA) patients, before and after surgical procedure, in order to assess the effect of tumor resection in the immune system homeostasis and to determine possible prognostic factors associated with high-grade tumors. We developed a whole-blood assay to monitor immune alterations and functional competence of peripheral monocytes in a group of 10 healthy individuals (HG), in 20 HCC patients and 8 CCA patients, by multi-color flow cytometry, qRT-PCR, and ELISA techniques. The qRT-PCR analysis showed an upregulation of TNFα expression by classical and intermediate monocytes purified from HCC patients presenting tumors in grade G3-G4 as compared to G1-G2 HCC patients. Moreover, ELISA assay confirmed elevated serum levels of TNFα in G3-G4 compared to G1-G2 HCC patients. A significant decrease of circulating non-classical monocytes was detected in both CCA and HCC patients before and after surgical procedure. In addition, a functional defect in circulating classical and intermediate monocytes was observed in both groups of cancer patients when compared to the HG, with partial recovery after the surgical intervention. This integrated analysis permitted the identification of altered functional competence of monocyte subsets in CCA and HCC patients. In addition, our results point to a potential role of TNFα as a prognostic peripheral biomarker in HCC patients, indicating the presence of high-grade tumors that should be further validated.

中文翻译：

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与高级别肝细胞癌相关的纯化外周血单核细胞亚群中可溶性 TNFα 水平升高和 TNFα mRNA 表达上调

慢性炎症涉及多种癌症的发生和进展，包括肝癌。目前的研究重点是肝细胞癌（HCC）和胆管癌（CCA）患者手术前后外周免疫反应的特征，以评估肿瘤切除对免疫系统稳态的影响并确定可能的预后与高级别肿瘤相关的因素。我们开发了一种全血测定法，通过多色流式细胞术、qRT-PCR 和 ELISA 监测 10 名健康个体 (HG)、20 名 HCC 患者和 8 名 CCA 患者的外周单核细胞的免疫变化和功能能力技术。qRT-PCR 分析显示，与 G1-G2 HCC 患者相比，从呈现 G3-G4 级肿瘤的 HCC 患者中纯化的经典单核细胞和中间单核细胞的 TNFα 表达上调。此外，ELISA 测定证实，与 G1-G2 HCC 患者相比，G3-G4 患者的血清 TNFα 水平升高。CCA 和 HCC 患者在手术前后均检测到循环非经典单核细胞显着减少。此外，与 HG 相比，两组癌症患者均观察到循环经典单核细胞和中间单核细胞的功能缺陷，且在手术干预后部分恢复。这种综合分析可以识别 CCA 和 HCC 患者单核细胞亚群功能能力的改变。此外，我们的结果指出 TNFα 作为 HCC 患者预后外周生物标志物的潜在作用，表明存在应进一步验证的高级别肿瘤。