TSG6 affects anti cancer drug resistance and angiogenesis in 3D spheroid model of canine mammary gland tumor cells

Abstract 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 microenvi...

Full description

Saved in:
Bibliographic Details
Main Authors: Ga-Hyun Lim, Ju-Hyun An, Jeong-Hwa Lee, Kyoung-Won Seo, Hwa-Young Youn
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Scientific Reports
Subjects:
Online Access:https://doi.org/10.1038/s41598-025-12950-3
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849226349296746496
author Ga-Hyun Lim
Ju-Hyun An
Jeong-Hwa Lee
Kyoung-Won Seo
Hwa-Young Youn
author_facet Ga-Hyun Lim
Ju-Hyun An
Jeong-Hwa Lee
Kyoung-Won Seo
Hwa-Young Youn
author_sort Ga-Hyun Lim
collection DOAJ
description Abstract 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.
format Article
id doaj-art-52ae2ae743654bd1ad49fa1fcbb3d8a9
institution Kabale University
issn 2045-2322
language English
publishDate 2025-08-01
publisher Nature Portfolio
record_format Article
series Scientific Reports
spelling doaj-art-52ae2ae743654bd1ad49fa1fcbb3d8a92025-08-24T11:22:07ZengNature PortfolioScientific Reports2045-23222025-08-0115111310.1038/s41598-025-12950-3TSG6 affects anti cancer drug resistance and angiogenesis in 3D spheroid model of canine mammary gland tumor cellsGa-Hyun Lim0Ju-Hyun An1Jeong-Hwa Lee2Kyoung-Won Seo3Hwa-Young Youn4Laboratory of Veterinary Internal Medicine, Department of Veterinary Clinical Science, College of Veterinary Medicine, Seoul National UniversityDepartment of Veterinary Emergency and Critical Care Medicine, Institute of Veterinary Science, College of Veterinary Medicine, Kangwon National UniversityK-BIO KIURI Center, Seoul National UniversityLaboratory of Veterinary Internal Medicine, Department of Veterinary Clinical Science, College of Veterinary Medicine, Seoul National UniversityLaboratory of Veterinary Internal Medicine, Department of Veterinary Clinical Science, College of Veterinary Medicine, Seoul National UniversityAbstract 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.https://doi.org/10.1038/s41598-025-12950-3AngiogenesisAnti-cancer drug resistanceCanine mammary gland tumorTSG6Tumor microenvironmentSpheroid
spellingShingle Ga-Hyun Lim
Ju-Hyun An
Jeong-Hwa Lee
Kyoung-Won Seo
Hwa-Young Youn
TSG6 affects anti cancer drug resistance and angiogenesis in 3D spheroid model of canine mammary gland tumor cells
Scientific Reports
Angiogenesis
Anti-cancer drug resistance
Canine mammary gland tumor
TSG6
Tumor microenvironment
Spheroid
title TSG6 affects anti cancer drug resistance and angiogenesis in 3D spheroid model of canine mammary gland tumor cells
title_full TSG6 affects anti cancer drug resistance and angiogenesis in 3D spheroid model of canine mammary gland tumor cells
title_fullStr TSG6 affects anti cancer drug resistance and angiogenesis in 3D spheroid model of canine mammary gland tumor cells
title_full_unstemmed TSG6 affects anti cancer drug resistance and angiogenesis in 3D spheroid model of canine mammary gland tumor cells
title_short TSG6 affects anti cancer drug resistance and angiogenesis in 3D spheroid model of canine mammary gland tumor cells
title_sort tsg6 affects anti cancer drug resistance and angiogenesis in 3d spheroid model of canine mammary gland tumor cells
topic Angiogenesis
Anti-cancer drug resistance
Canine mammary gland tumor
TSG6
Tumor microenvironment
Spheroid
url https://doi.org/10.1038/s41598-025-12950-3
work_keys_str_mv AT gahyunlim tsg6affectsanticancerdrugresistanceandangiogenesisin3dspheroidmodelofcaninemammaryglandtumorcells
AT juhyunan tsg6affectsanticancerdrugresistanceandangiogenesisin3dspheroidmodelofcaninemammaryglandtumorcells
AT jeonghwalee tsg6affectsanticancerdrugresistanceandangiogenesisin3dspheroidmodelofcaninemammaryglandtumorcells
AT kyoungwonseo tsg6affectsanticancerdrugresistanceandangiogenesisin3dspheroidmodelofcaninemammaryglandtumorcells
AT hwayoungyoun tsg6affectsanticancerdrugresistanceandangiogenesisin3dspheroidmodelofcaninemammaryglandtumorcells