HBV 介导的 S100A9-TLR4/RAGE-ROS 级联反应导致中性粒细胞胞外诱捕网升高，促进肝癌的生长和转移。

Elevated neutrophil extracellular traps by HBV-mediated S100A9-TLR4/RAGE-ROS cascade facilitate the growth and metastasis of hepatocellular carcinoma.

机构信息

Department of Laboratory Medicine, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400000, P. R. China.

Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, P. R. China.

出版信息

Cancer Commun (Lond). 2023 Feb;43(2):225-245. doi: 10.1002/cac2.12388. Epub 2022 Nov 8.

DOI:10.1002/cac2.12388

PMID:36346061

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9926958/

Abstract

BACKGROUND

Neutrophil extracellular traps (NETs) are considered significant contributors to cancer progression, especially metastasis. However, it is still unclear whether NETs are involved in hepatitis B virus (HBV)-related hepatocarcinogenesis and have potential clinical significance during evaluation and management for hepatocellular carcinoma (HCC). In this study, we aimed to investigate the functional mechanism of NETs in HBV-related hepatocarcinogenesis and their clinical significance.

METHODS

A total of 175 HCC patients with and without HBV infection and 58 healthy controls were enrolled in this study. NETs were measured in tissue specimens, freshly isolated neutrophils and blood serum from these patients, and the correlation of circulating serum NETs levels with malignancy was evaluated. The mechanism by which HBV modulates NETs formation was explored using cell-based studies. In addition, in vitro and in vivo experiments were further performed to clarify the functional mechanism of NETs on the growth and metastasis of HCC.

RESULTS

We observed an elevated level of NETs in blood serum and tissue specimens from HCC patients, especially those infected with HBV. NETs facilitated the growth and metastasis of HCC both in vitro and in vivo, which were mainly dominated by increased angiogenesis, epithelial-mesenchymal transition (EMT)-related cell migration, matrix metalloproteinases (MMPs)-induced extracellular matrix (ECM) degradation and NETs-mediated cell trapping. Inhibition of NETs generation by DNase 1 effectively abrogated the NETs-aroused HCC growth and metastasis. In addition, HBV-induced S100A9 accelerated the generation of NETs, which was mediated by activation of toll-like receptor (TLR4)/receptor for advanced glycation end products (RAGE)-reactive oxygen species (ROS) signaling. Further, circulatory NETs were found to correlate with viral load, TNM stage and metastasis status in HBV-related HCC, and the identified NETs could predict extrahepatic metastasis, with an area under the ROC curve (AUC) of 0.83 and 90.3% sensitivity and 62.8% specificity at a cutoff value of 0.32.

CONCLUSIONS

Our findings indicated that activation of RAGE/TLR4-ROS signaling by HBV-induced S100A9 resulted in abundant NETs formation, which subsequently facilitated the growth and metastasis of HCC cells. More importantly, the identified circulatory NETs exhibited potential as an alternative biomarker for predicting extrahepatic metastasis in HBV-related HCC.

摘要

背景

中性粒细胞胞外诱捕网（NETs）被认为是癌症进展的重要因素，尤其是转移。然而，目前尚不清楚 NETs 是否参与乙型肝炎病毒（HBV）相关的肝癌发生，以及它们在肝细胞癌（HCC）评估和管理中的潜在临床意义。在本研究中，我们旨在研究 NETs 在 HBV 相关肝癌发生中的功能机制及其临床意义。

方法

本研究共纳入了 175 例 HBV 感染和未感染的 HCC 患者以及 58 例健康对照者。检测了这些患者组织标本、新鲜分离的中性粒细胞和血清中的 NETs，并评估了循环血清 NETs 水平与恶性肿瘤的相关性。通过细胞基础研究探索了 HBV 调节 NETs 形成的机制。此外，还进行了体外和体内实验，以阐明 NETs 对 HCC 生长和转移的功能机制。

结果

我们观察到 HCC 患者的血清和组织标本中 NETs 水平升高，尤其是感染 HBV 的患者。NETs 促进了 HCC 在体外和体内的生长和转移，主要由血管生成增加、上皮-间充质转化（EMT）相关细胞迁移、基质金属蛋白酶（MMPs）诱导的细胞外基质（ECM）降解和 NETs 介导的细胞捕获所介导。DNase 1 抑制 NETs 的生成可有效阻断 NETs 引起的 HCC 生长和转移。此外，HBV 诱导的 S100A9 通过激活 toll 样受体（TLR4）/晚期糖基化终产物受体（RAGE）-活性氧（ROS）信号转导加速 NETs 的生成。进一步研究发现，HBV 相关 HCC 患者的循环 NETs 与病毒载量、TNM 分期和转移状态相关，并且鉴定出的 NETs 可以预测肝外转移，ROC 曲线下面积（AUC）为 0.83，cutoff 值为 0.32 时，敏感性为 90.3%，特异性为 62.8%。