Endoscopy Center, Minhang Hospital, Fudan University, Shanghai, 201199, China.
J Cancer Res Clin Oncol. 2023 Aug;149(10):7053-7067. doi: 10.1007/s00432-023-04655-9. Epub 2023 Mar 2.
Tumor-associated macrophages (TAMs) constitute the main infiltrating immune cells in the solid tumor microenvironment. Amounting studies have analyzed the antitumor effect on immune response induced by Toll-like receptor (TLR) agonists, such as lipopolysaccharide (LPS), γ-interferon (γ-IFN), and palmitic Acid (PA). However, their combined treatment for gastric cancer (GC) has not been illuminated.
We investigated the relevance of macrophage polarization and the effect of PA and γ-IFN in GC in vitro and in vivo. M1 and M2 macrophage-associated markers were measured by real-time quantitative PCR and flow cytometry, and the activation level of the TLR4 signaling pathways was evaluated by western blot analysis. The effect of PA and γ-IFN on the proliferation, migration, and invasion of GC cells (GCCs) was evaluated by Cell-Counting Kit-8, transwell assays, and wound-healing assays. In vivo animal models were used to verify the effect of PA and γ-IFN on tumor progression, and the M1 and M2 macrophage markers, CD8 + T lymphocytes, regulatory T cells (Treg) cells, and the myeloid-derived suppressor cells (MDSCs) in tumor tissues were analyzed by flow cytometry and immunohistochemical (IHC).
The results showed that this combination strategy enhanced M1-like macrophages and diminished M2-like macrophages through the TLR4 signaling pathway in vitro. In addition, the combination strategy impairs the proliferative and migratory activity of GCC in vitro and in vivo. While, the antitumor effect was abolished using the TAK-424 (a specific TLR-4 signaling pathway inhibitor) in vitro.
The combined treatment of PA and γ-IFN inhibited GC progression by modulating macrophages polarization via the TLR4 pathway.
肿瘤相关巨噬细胞（TAMs）构成实体瘤微环境中主要浸润的免疫细胞。大量研究分析了 Toll 样受体（TLR）激动剂（如脂多糖（LPS）、γ-干扰素（γ-IFN）和棕榈酸（PA））诱导的免疫反应的抗肿瘤作用。然而，它们在胃癌（GC）中的联合治疗尚未阐明。
我们在体外和体内研究了巨噬细胞极化和 PA 和 γ-IFN 在 GC 中的相关性及其作用。通过实时定量 PCR 和流式细胞术测量 M1 和 M2 巨噬细胞相关标志物，并通过 Western blot 分析评估 TLR4 信号通路的激活水平。通过细胞计数试剂盒-8（Cell-Counting Kit-8）、Transwell 测定和划痕愈合测定评估 PA 和 γ-IFN 对 GC 细胞（GCCs）增殖、迁移和侵袭的影响。体内动物模型用于验证 PA 和 γ-IFN 对肿瘤进展的影响，并通过流式细胞术和免疫组织化学（IHC）分析肿瘤组织中的 M1 和 M2 巨噬细胞标志物、CD8+T 淋巴细胞、调节性 T 细胞（Treg）细胞和髓源性抑制细胞（MDSCs）。
结果表明，这种联合策略通过 TLR4 信号通路在体外增强了 M1 样巨噬细胞并减少了 M2 样巨噬细胞。此外，联合策略损害了 GCC 在体外和体内的增殖和迁移活性。然而，在体外使用 TAK-424（一种特异性 TLR-4 信号通路抑制剂）时，这种抗肿瘤作用被消除。
PA 和 γ-IFN 的联合治疗通过 TLR4 通路调节巨噬细胞极化抑制 GC 进展。
