Lim Ga-Hyun, An Ju-Hyun, Lee Jeong-Hwa, Seo Kyoung-Won, Youn Hwa-Young
Laboratory of Veterinary Internal Medicine, Department of Veterinary Clinical Science, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
VIP animal medical center, Seoul, Republic of Korea.
Sci Rep. 2025 Aug 22;15(1):30852. doi: 10.1038/s41598-025-12950-3.
TSG6 (Tumor necrosis factor stimulated gene-6) plays a critical role in modulating the tumor microenvironment by regulating inflammation, immune responses, and extracellular matrix remodeling. Hypoxia affects tumor growth, angiogenesis, and anti-cancer drug resistance in the tumor microenvironment, and TSG6 expression is known to influence HIF-1α expression in tumor tissues. In this study, we created TSG6 knockdown spheroids of canine mammary gland tumor (MGT) cells (CIPp and CIPm) to investigate the effects of TSG6 on angiogenesis and drug resistance in the tumor microenvironment. Using a siRNA transfection system, we induced TSG6 knockdown in canine MGT cells and formed TSG6 knockdown spheroids using an ultra-low adhesion plate. Cell viability and apoptosis were assessed by treating TSG6 knockdown spheroids with doxorubicin. Angiogenesis was evaluated by forming vascularized spheroids with canine MGT and endothelial cells (ECs). TSG6 knockdown led to reduced expression of tumor growth factors and multidrug resistance genes in canine MGT cells, as well as a significant reduction in hypoxic conditions within the spheroids. When treated with doxorubicin, TSG6 knockdown spheroids exhibited decreased viability and increased apoptosis. In a vascularized TSG6 knockdown spheroid model, TSG6 knockdown significantly reduced the expression of CD31 and tube formation in canine ECs. In conclusion, we created a TSG6 knockdown spheroid model to investigate the role of TSG6 in the tumor microenvironment. TSG6 knockdown significantly reduced anti-cancer drug resistance and angiogenesis in canine MGT cells. Therefore, TSG6 could be considered a potential therapeutic target for canine MGT.
TSG6（肿瘤坏死因子刺激基因-6）通过调节炎症、免疫反应和细胞外基质重塑，在调节肿瘤微环境中发挥关键作用。缺氧影响肿瘤微环境中的肿瘤生长、血管生成和抗癌药物耐药性，并且已知TSG6表达会影响肿瘤组织中HIF-1α的表达。在本研究中，我们构建了犬乳腺肿瘤（MGT）细胞（CIPp和CIPm）的TSG6敲低球体，以研究TSG6对肿瘤微环境中血管生成和耐药性的影响。使用siRNA转染系统，我们在犬MGT细胞中诱导TSG6敲低，并使用超低附着板形成TSG6敲低球体。通过用阿霉素处理TSG6敲低球体来评估细胞活力和凋亡。通过将犬MGT细胞与内皮细胞（ECs）形成血管化球体来评估血管生成。TSG6敲低导致犬MGT细胞中肿瘤生长因子和多药耐药基因的表达降低，以及球体中缺氧条件的显著减少。当用阿霉素处理时，TSG6敲低球体表现出活力降低和凋亡增加。在血管化的TSG6敲低球体模型中，TSG6敲低显著降低了犬ECs中CD31的表达和管形成。总之，我们构建了一个TSG6敲低球体模型来研究TSG6在肿瘤微环境中的作用。TSG6敲低显著降低了犬MGT细胞中的抗癌药物耐药性和血管生成。因此，TSG6可被认为是犬MGT的一个潜在治疗靶点。
