Ishihara Nobuaki, Koma Yu-Ichiro, Omori Masaki, Komatsu Shohei, Torigoe Rikuya, Yokoo Hiroki, Nakanishi Takashi, Yamanaka Keitaro, Azumi Yuki, Tsukamoto Shuichi, Kodama Takayuki, Nishio Mari, Shigeoka Manabu, Yokozaki Hiroshi, Fukumoto Takumi
Division of Pathology, Department of Pathology, Kobe University Graduate School of Medicine, Kobe, Japan; Division of Hepato-Biliary-Pancreatic Surgery, Department of Surgery, Kobe University Graduate School of Medicine, Kobe, Japan.
Division of Pathology, Department of Pathology, Kobe University Graduate School of Medicine, Kobe, Japan.
Am J Pathol. 2025 Mar;195(3):589-608. doi: 10.1016/j.ajpath.2024.12.007. Epub 2025 Jan 3.
Tumor-infiltrating macrophages, known as tumor-associated macrophages, play a crucial role in the tumor microenvironment. Herein, immunohistochemistry revealed that intratumoral CD68-positive macrophages are associated with poor prognosis and clinicopathologic factors in patients with hepatocellular carcinoma (HCC). Subsequently, an indirect co-culture system involving HCC cells and peripheral blood-derived macrophages was developed. cDNA microarray analysis revealed that chemokine (C-C motif) ligand 2 (CCL2) was highly expressed in HCC cells co-cultured with macrophages. CCL2 neutralization suppressed proliferation, migration, and phosphorylation of extracellular signal-regulated kinase (Erk) in HCC cells and macrophages enhanced through co-culture. In contrast, recombinant human CCL2 (rhCCL2) addition facilitated these malignant phenotypes and increased Erk phosphorylation levels in HCC cells and macrophages. The primary CCL2 receptor, CCR2, was expressed in HCC cells and macrophages and was up-regulated in co-cultured HCC cells. CCR2 inhibition suppressed malignant phenotypes and reduced phosphorylated levels of Erk enhanced by rhCCL2. Additionally, the inhibition of Erk signal suppressed rhCCL2-enhanced malignant phenotypes. Moreover, serum CCL2 levels were higher in patients with HCC than those in healthy donors. On the basis of immunohistochemistry, CCL2-positive cases with high CCR2 expression and phosphorylated Erk-positive cases exhibited poor survival outcomes. Therefore, CCL2 up-regulation through interactions between HCC cells and macrophages contributed to HCC progression, making the CCL2/CCR2/Erk signal a potential target for HCC treatment.
肿瘤浸润巨噬细胞，即所谓的肿瘤相关巨噬细胞，在肿瘤微环境中起着关键作用。在此，免疫组织化学显示，肿瘤内CD68阳性巨噬细胞与肝细胞癌（HCC）患者的预后不良及临床病理因素相关。随后，建立了一种涉及肝癌细胞和外周血来源巨噬细胞的间接共培养系统。cDNA微阵列分析显示，趋化因子（C-C基序）配体2（CCL2）在与巨噬细胞共培养的肝癌细胞中高表达。CCL2中和可抑制肝癌细胞的增殖、迁移以及细胞外信号调节激酶（Erk）的磷酸化，而通过共培养增强的巨噬细胞中Erk磷酸化水平也受到抑制。相反，添加重组人CCL2（rhCCL2）则促进了这些恶性表型，并增加了肝癌细胞和巨噬细胞中Erk的磷酸化水平。CCL2的主要受体CCR2在肝癌细胞和巨噬细胞中表达，并在共培养的肝癌细胞中上调。CCR2抑制可抑制恶性表型，并降低rhCCL2增强的Erk磷酸化水平。此外，抑制Erk信号可抑制rhCCL2增强的恶性表型。而且，HCC患者血清CCL2水平高于健康供体。基于免疫组织化学，CCL2高表达且CCR2阳性以及磷酸化Erk阳性的病例生存结局较差。因此，肝癌细胞与巨噬细胞相互作用导致的CCL2上调促进了肝癌进展，使得CCL2/CCR2/Erk信号成为肝癌治疗的潜在靶点。
