Li Cong, Zhang Hongan, Liu Yanchun, Zhang Ting, Gu Feng
Hepatobiliary Department, Hebei University Affiliated Hospital, Baoding, China.
Department of Pediatrics, North China Petroleum Administration General Hospital, Renqiu, China.
Sci Rep. 2025 May 29;15(1):18820. doi: 10.1038/s41598-025-02447-4.
To delineate Gpr109A's role and mechanisms in modulating the immune microenvironment of hepatocellular carcinoma. Employing Gpr109A-knockout mice and in vitro co-cultures of hepatocellular carcinoma cells with macrophages, this study utilized a suite of techniques, including lentiviral vectors for stable cell line establishment, Western blotting, cell scratch, CCK-8, transwell assays, flow cytometry, immunohistochemistry and phagocytosis assay to assess various cellular behaviors and interactions. Gpr109A deletion markedly reduced the oncogenic potential of H22 cells, both in vivo and when co-cultured with knockout macrophages, impairing their growth, invasion, and migration. In Gpr109A-knockout macrophages, an upregulation of MerTK and a reduction in immunosuppressive cytokine release were observed, indicating a shift towards an M2c macrophage phenotype. This shift is linked to Gpr109A's role in promoting protease overexpression and inhibiting SHP2 phosphorylation, crucial for enhancing cancer cell proliferation and invasiveness. Gpr109A significantly influences macrophage polarization to the M2c type, augmenting hepatocellular carcinoma cell aggressiveness.
为了阐明Gpr109A在调节肝细胞癌免疫微环境中的作用和机制。本研究采用Gpr109A基因敲除小鼠以及肝癌细胞与巨噬细胞的体外共培养体系，运用了一系列技术，包括用于建立稳定细胞系的慢病毒载体、蛋白质免疫印迹法、细胞划痕实验、CCK-8法、Transwell实验、流式细胞术、免疫组织化学和吞噬实验，以评估各种细胞行为和相互作用。Gpr109A基因缺失显著降低了H22细胞在体内以及与基因敲除巨噬细胞共培养时的致癌潜能，损害了它们的生长、侵袭和迁移能力。在Gpr109A基因敲除的巨噬细胞中，观察到MerTK的上调和免疫抑制细胞因子释放的减少，表明向M2c巨噬细胞表型转变。这种转变与Gpr109A在促进蛋白酶过表达和抑制SHP2磷酸化方面的作用有关，这对增强癌细胞增殖和侵袭至关重要。Gpr109A显著影响巨噬细胞向M2c型极化，增强肝癌细胞的侵袭性。
