Department of Oncology, Innovation centre for Advanced Interdisciplinary Medicine, Guangzhou Key Laboratory of Enhanced Recovery after Abdominal Surgery, The Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou Medical University, Guangzhou, 510700, China.
State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou Medical University, Guangzhou, 510062, China.
Drug Deliv. 2022 Dec;29(1):1712-1725. doi: 10.1080/10717544.2022.2079769.
This study aimed to explore the anti-tumor effect of icaritin loading poly (lactic-co-glycolic acid) nanoparticles (refer to PLGA@Icaritin NPs) on gastric cancer (GC) cells. Transmission Electron Microscope (TEM), size distribution, zeta potential, drug-loading capability, and other physicochemical characteristics of PLGA@Icaritin NPs were carried out. Furthermore, flow cytometry, confocal laser scanning microscope (CLSM), Cell Counting Kit-8 (CCK-8), Transwell, Elisa assay and Balb/c mice were applied to explore the cellular uptake, anti-proliferation, anti-metastasis, immune response activation effects, and related anti-tumor mechanism of PLGA@Icaritin NPs and . PLGA@Icaritin NPs showed spherical shape, with appropriate particle sizes and well drug loading and releasing capacities. Flow cytometry and CLSM results indicated that PLGA@Icaritin could efficiently enter into GC cells. CCK-8 proved that PLGA@Icaritin NPs dramatically suppressed cell growth, induced Lactic dehydrogenase (LDH) leakage, arrested more GC cells at G2 phase, and inhibited the invasion and metastasis of GC cells, compared to free icaritin. In addition, PLGA@Icaritin could help generate dozens of reactive oxygen species (ROS) within GC cells, following by significant mitochondrial membrane potentials (MMPs) loss and excessive production of oxidative-mitochondrial DNA (Ox-mitoDNA). Since that, Ox-mitoDNA further activated the releasing of damage associated molecular pattern molecules (DAMPs), and finally led to immunogenic cell death (ICD). Our data also elaborated that PLGA@Icaritin exerted a powerful inhibitory effect (∼80%), compared to free icaritin (∼60%). Most importantly, our results demonstrated that PLGA@Icaritin could activate the anti-tumor immunity via recruitment of infiltrating CD4+ cells, CD8+ T cells and increased secretion of cytokine immune factors, including interferon-γ (IFN-γ) tumor necrosis factor-α (TNF-α) and interleukin-1 (IL-1). Our findings validate that the successful design of PLGA@Icaritin, which can effectively active ICD and facilitate tumor recruitment in GC through inducing mitoDNA oxidative damage.
本研究旨在探索淫羊藿次苷 PLGA@Icaritin NPs 对胃癌（GC）细胞的抗肿瘤作用。透射电子显微镜（TEM）、粒径分布、Zeta 电位、载药能力等 PLGA@Icaritin NPs 的理化特性进行了研究。此外，流式细胞术、共聚焦激光扫描显微镜（CLSM）、细胞计数试剂盒-8（CCK-8）、Transwell、酶联免疫吸附试验（ELISA）和 Balb/c 小鼠被用于探讨 PLGA@Icaritin NPs 的细胞摄取、抗增殖、抗转移、免疫反应激活作用及相关抗肿瘤机制。PLGA@Icaritin NPs 呈球形，具有合适的粒径和良好的载药和释放能力。流式细胞术和 CLSM 结果表明，PLGA@Icaritin 能够有效地进入 GC 细胞。CCK-8 证明 PLGA@Icaritin NPs 能显著抑制细胞生长，诱导乳酸脱氢酶（LDH）漏出，使更多的 GC 细胞停滞在 G2 期，并抑制 GC 细胞的侵袭和转移，与游离淫羊藿素相比。此外，PLGA@Icaritin 能在 GC 细胞内产生数十种活性氧（ROS），随后导致线粒体膜电位（MMPs）显著丧失和氧化线粒体 DNA（Ox-mitoDNA）的过度产生。因此，Ox-mitoDNA 进一步激活了损伤相关分子模式分子（DAMPs）的释放，最终导致免疫原性细胞死亡（ICD）。我们的数据还表明，与游离淫羊藿素（约 60%）相比，PLGA@Icaritin 发挥了强大的抑制作用（约 80%）。最重要的是，我们的结果表明，PLGA@Icaritin 能够通过募集浸润性 CD4+细胞、CD8+T 细胞和增加细胞因子免疫因子的分泌来激活抗肿瘤免疫，包括干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）和白细胞介素-1（IL-1）。我们的研究结果证实了 PLGA@Icaritin 的成功设计，它可以通过诱导 mitoDNA 氧化损伤，有效地激活 ICD，并促进 GC 中的肿瘤募集。
