Neutrophil Extracellular Traps Promote Hepatocellular Carcinoma Cell Proliferation and Impair Hepatocyte Function by Remodeling the Extracellular Matrix
ZHONG Hongling1#, WANG Jinli2#, XIAO Sijia2, SHEN Kan1, QIAN Chen1, WANG Wenxi2, ZHAN Shuyu1, CHENG Shuqun3, ZHENG Yongxia1*
HCC (hepatocellular carcinoma) is a prevalent malignant tumor, and chronic inflammation is a critical driving force in its development. NETs (neutrophil extracellular traps), as key products of the inflammatory response, play a significant role in the tumor microenvironment. This study aimed to investigate the role of NETs in regulating HCC proliferation and metabolic reprogramming. In vitro, a co-culture system of NETs and Hep3B was established. The effects of NETs on HCC cell proliferation were evaluated using CCK-8 assays, cell cycle analysis, and soft agar colony formation assays. In vivo, a subcutaneous tumor xenograft model was established in mice, and NETs were induced by lipopolysaccharide. Tumor growth was evaluated by measuring tumor size and examining liver pathological changes. Moreover, the effects of NETs on the metabolism and function of liver cancer cells were explored by detecting glycogen content in liver cancer tissues and cells and the expression of glycolysis-related genes, respectively. Finally, immunofluorescence staining was used to detect the expression of Fn (fibronectin) in the extracellular matrix to explore the underlying mechanisms, and Western blot was used to measure the activity of the integrin pathway mediated by Fn. Results showed that co-culture with NETs promoted the proliferation of Hep3B cells in vitro. In vivo, NETs promoted tumor growth and inflammatory cell infiltration in the liver of C57BL/6 mice. Additionally, NETs decreased glycogen levels in HCC cells, enhanced glycolytic activity, and increased the levels of ALT (alanine transaminase) in the cell culture supernatant, suggesting impaired liver function. Mechanistically, NETs increased the expression of Fn in the extracellular matrix, remodeling the tumor microenvironment and providing a favorable condition for tumor cell growth. Therefore, this study reveals that NETs promote HCC proliferation by inducing ECM remodeling, enhance glycolysis, and impair hepatocyte function.
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