Long-term resident adipose-derived stromal stem cells in the microenvironment remodeling BLCA cell stemness and EMT promotes bladder cancer progression

Abstract The role of adipose-derived stromal stem cells (ADSCs) in BLCA progression is unclear. We investigated the effects of invasion, stemness, Epithelial-mesenchymal transition (EMT), and drug resistance of BLCA cells co-cultured with ADSCs for a long period of time. Cells were divided into six...

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Main Authors: Denggao Huang, Na Guo, Yanling Peng, Zhenyu Nie, Yuanhui Gao, Hui Cao, Shunlan Wang, Mei Chen, Xiaohong Wen, Zhaojun Wang, Linlin Zheng, Haowei He, Yong Sun, Yang Chen, Shufang Zhang
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Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-07387-7
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author Denggao Huang
Na Guo
Yanling Peng
Zhenyu Nie
Yuanhui Gao
Hui Cao
Shunlan Wang
Mei Chen
Xiaohong Wen
Zhaojun Wang
Linlin Zheng
Haowei He
Yong Sun
Yang Chen
Shufang Zhang
author_facet Denggao Huang
Na Guo
Yanling Peng
Zhenyu Nie
Yuanhui Gao
Hui Cao
Shunlan Wang
Mei Chen
Xiaohong Wen
Zhaojun Wang
Linlin Zheng
Haowei He
Yong Sun
Yang Chen
Shufang Zhang
author_sort Denggao Huang
collection DOAJ
description Abstract The role of adipose-derived stromal stem cells (ADSCs) in BLCA progression is unclear. We investigated the effects of invasion, stemness, Epithelial-mesenchymal transition (EMT), and drug resistance of BLCA cells co-cultured with ADSCs for a long period of time. Cells were divided into six groups: ADSCs group, ADSCs: T24 group (10:1, 3:1 and 1:1 groups), ADSCs-derived conditioned medium group (CM) and T24 cell group (T24), and cells in each group were cultured to 14 days, and puromycin (puro) was added to the co-cultured cell to remove ADSCs cells without puro resistance, and then the function of T24 cells (10:1-COC, 3:1-COC and 1:1-COC) after co-culture was studied; CCK-8 assay, Transwell, Wound healing, Flow cytometry, RNA-sequencing, qRT-PCR and Western Bloting assay were used to detect cell proliferation, invasion, migration, apoptosis and cellular mRNA and protein expression levels, respectively. We unexpectedly found enhanced stemness and drug resistance of BLCA cells after prolonged contact culture with ADSCs. T24 cells after co-culture mediated cell proliferation, invasion, EMT, stemness, drug resistance and immune escape by up-regulating MDM2, mt-P53 and PD-L1, compared to CM and T24 groups. The inhibitor Atezo and CP-31,398 eliminated mt-P53 and PD-L1-mediated T24 cell drug resistance and stemness, respectively. This study demonstrated that after prolonged co-culture of BLCA cells with ADSCs, the stemness, drug resistance, and immune evasion of T24 cells were dramatically enhanced, suggesting that long-term resident ADSCs in the bladder cancer tumor microenvironment play a procarcinogenic role.
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spelling doaj-art-e3661d78eb7d47ad8b4c6c5d678e284c2025-08-20T03:45:20ZengNature PortfolioScientific Reports2045-23222025-07-0115111510.1038/s41598-025-07387-7Long-term resident adipose-derived stromal stem cells in the microenvironment remodeling BLCA cell stemness and EMT promotes bladder cancer progressionDenggao Huang0Na Guo1Yanling Peng2Zhenyu Nie3Yuanhui Gao4Hui Cao5Shunlan Wang6Mei Chen7Xiaohong Wen8Zhaojun Wang9Linlin Zheng10Haowei He11Yong Sun12Yang Chen13Shufang Zhang14Central Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalCentral Laboratory, Haikou Affiliated Hospital of Central South University Xiangya School of MedicalAbstract The role of adipose-derived stromal stem cells (ADSCs) in BLCA progression is unclear. We investigated the effects of invasion, stemness, Epithelial-mesenchymal transition (EMT), and drug resistance of BLCA cells co-cultured with ADSCs for a long period of time. Cells were divided into six groups: ADSCs group, ADSCs: T24 group (10:1, 3:1 and 1:1 groups), ADSCs-derived conditioned medium group (CM) and T24 cell group (T24), and cells in each group were cultured to 14 days, and puromycin (puro) was added to the co-cultured cell to remove ADSCs cells without puro resistance, and then the function of T24 cells (10:1-COC, 3:1-COC and 1:1-COC) after co-culture was studied; CCK-8 assay, Transwell, Wound healing, Flow cytometry, RNA-sequencing, qRT-PCR and Western Bloting assay were used to detect cell proliferation, invasion, migration, apoptosis and cellular mRNA and protein expression levels, respectively. We unexpectedly found enhanced stemness and drug resistance of BLCA cells after prolonged contact culture with ADSCs. T24 cells after co-culture mediated cell proliferation, invasion, EMT, stemness, drug resistance and immune escape by up-regulating MDM2, mt-P53 and PD-L1, compared to CM and T24 groups. The inhibitor Atezo and CP-31,398 eliminated mt-P53 and PD-L1-mediated T24 cell drug resistance and stemness, respectively. This study demonstrated that after prolonged co-culture of BLCA cells with ADSCs, the stemness, drug resistance, and immune evasion of T24 cells were dramatically enhanced, suggesting that long-term resident ADSCs in the bladder cancer tumor microenvironment play a procarcinogenic role.https://doi.org/10.1038/s41598-025-07387-7Adipose-derived stromal stem cellsBladder cancerStemnessEpithelial mesenchymal transitionInvasionDrug resistance
spellingShingle Denggao Huang
Na Guo
Yanling Peng
Zhenyu Nie
Yuanhui Gao
Hui Cao
Shunlan Wang
Mei Chen
Xiaohong Wen
Zhaojun Wang
Linlin Zheng
Haowei He
Yong Sun
Yang Chen
Shufang Zhang
Long-term resident adipose-derived stromal stem cells in the microenvironment remodeling BLCA cell stemness and EMT promotes bladder cancer progression
Scientific Reports
Adipose-derived stromal stem cells
Bladder cancer
Stemness
Epithelial mesenchymal transition
Invasion
Drug resistance
title Long-term resident adipose-derived stromal stem cells in the microenvironment remodeling BLCA cell stemness and EMT promotes bladder cancer progression
title_full Long-term resident adipose-derived stromal stem cells in the microenvironment remodeling BLCA cell stemness and EMT promotes bladder cancer progression
title_fullStr Long-term resident adipose-derived stromal stem cells in the microenvironment remodeling BLCA cell stemness and EMT promotes bladder cancer progression
title_full_unstemmed Long-term resident adipose-derived stromal stem cells in the microenvironment remodeling BLCA cell stemness and EMT promotes bladder cancer progression
title_short Long-term resident adipose-derived stromal stem cells in the microenvironment remodeling BLCA cell stemness and EMT promotes bladder cancer progression
title_sort long term resident adipose derived stromal stem cells in the microenvironment remodeling blca cell stemness and emt promotes bladder cancer progression
topic Adipose-derived stromal stem cells
Bladder cancer
Stemness
Epithelial mesenchymal transition
Invasion
Drug resistance
url https://doi.org/10.1038/s41598-025-07387-7
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