Xie Yahui, Lv Zhen, Wang Yubin, Ma Jin, Wei Xingmin, Zheng Guisen, Wu Jianjun
School of Public Health, Gansu University of Traditional Chinese Medicine, Lanzhou, China.
Center for Laboratory Medicine, The Second Hospital of Lanzhou University, Lanzhou, China.
Front Immunol. 2025 Mar 13;16:1554467. doi: 10.3389/fimmu.2025.1554467. eCollection 2025.
Soluble programmed cell death receptor-1 (sPD-1) blocks the PD-1/PD-L1 pathway, reverses tumor immune suppression, and inhibits tumor growth. However, clinical applications are limited by its poor tissue distribution and rapid dispersion. Bone marrow-derived mesenchymal stem cells (BMSCs) are favorable carriers for tumor immunotherapy due to their capacity for external gene introduction and targeted tumor homing. However, they may inadvertently promote tumor growth. Interferon-gamma (IFN-γ) inhibits BMSC-mediated tumor growth and stimulates antigen-presenting cells to activate T lymphocytes. This study utilizes BMSCs transfected with IFN-γ as carriers for sPD-1, enabling the targeted homing of sPD-1 to tumor tissues, thereby enhancing the efficacy and sustained stability of immunotherapy.
stable IFN-γ- and sPD-1-overexpressing BMSCs were successfully constructed by lentiviral transfection. A non-contact co-culture system was established with Lewis and A549 lung adenocarcinoma cells to observe changes in the lung cancer cells after co-culture, using assays including cell migration and invasion experiments, as well as cellular senescence detection. Additionally, a subcutaneous lung adenocarcinoma model was established in C57BL/6J mice for intervention studies. Tumor volume, cellular apoptosis in tumor tissue (assessed by TUNEL assay), peripheral Treg cells (analyzed by flow cytometry), and histopathological markers (evaluated by HE staining and immunohistochemistry) were analyzed. The expression levels of BAX, BCL-2, AKT, PI3K, and PD-L1 were assessed by quantitative PCR and Western Blot.
IFN-γ- and sPD-1-overexpressing BMSCs exhibited high bioactivity and genetic stability, inhibiting lung adenocarcinoma cell proliferation, accelerating cellular senescence, and reducing migration and invasion. Furthermore, they upregulate Bax expression, downregulate Bcl-2, and promote apoptosis. Additionally, these cells alleviate inflammatory damage in lung tissue of tumor-bearing mice, lower Treg cell levels to inhibit tumor immune evasion, and reduce the expression of PI3K/AKT and PD-L1.
IFN-γ- and sPD-1-overexpressing BMSCs effectively inhibit lung adenocarcinoma cell growth and tumor progression. The primary mechanisms include suppression of cancer cell growth, migration, and invasion; promotion of apoptosis and senescence in cancer cells; modulation of Treg cells; and inhibition of the PI3K/AKT signaling pathway and PD-1/PD-L1 pathways.
可溶性程序性细胞死亡受体1（sPD-1）可阻断PD-1/PD-L1通路，逆转肿瘤免疫抑制并抑制肿瘤生长。然而，其临床应用受限于较差的组织分布和快速弥散。骨髓间充质干细胞（BMSCs）因其具有导入外源基因和靶向肿瘤归巢的能力，是肿瘤免疫治疗的理想载体。然而，它们可能会意外促进肿瘤生长。干扰素-γ（IFN-γ）可抑制BMSC介导的肿瘤生长，并刺激抗原呈递细胞激活T淋巴细胞。本研究利用转染了IFN-γ的BMSCs作为sPD-1的载体，使sPD-1靶向归巢至肿瘤组织，从而提高免疫治疗的疗效和持续稳定性。
通过慢病毒转染成功构建了稳定过表达IFN-γ和sPD-1的BMSCs。与Lewis和A549肺腺癌细胞建立非接触共培养系统，采用细胞迁移和侵袭实验以及细胞衰老检测等方法观察共培养后肺癌细胞的变化。此外，在C57BL/6J小鼠中建立皮下肺腺癌模型进行干预研究。分析肿瘤体积、肿瘤组织中的细胞凋亡（通过TUNEL检测评估）、外周调节性T细胞（通过流式细胞术分析）以及组织病理学标志物（通过HE染色和免疫组织化学评估）。通过定量PCR和蛋白质免疫印迹法评估BAX、BCL-2、AKT、PI3K和PD-L1的表达水平。
过表达IFN-γ和sPD-1的BMSCs表现出高生物活性和遗传稳定性，抑制肺腺癌细胞增殖，加速细胞衰老，并减少迁移和侵袭。此外，它们上调Bax表达，下调Bcl-2，并促进细胞凋亡。此外，这些细胞减轻荷瘤小鼠肺组织的炎症损伤，降低调节性T细胞水平以抑制肿瘤免疫逃逸，并降低PI3K/AKT和PD-L1的表达。
过表达IFN-γ和sPD-1的BMSCs有效抑制肺腺癌细胞生长和肿瘤进展。主要机制包括抑制癌细胞生长、迁移和侵袭；促进癌细胞凋亡和衰老；调节调节性T细胞；以及抑制PI3K/AKT信号通路和PD-1/PD-L1通路。
