Adaptive 3D Self‐Assembly of Colorectal Cancer Cells With Unchanged Tumor Phenotype and Drug Sensitivity

ABSTRACT Background Three‐dimensional (3D) self‐assembly of organoids or tumoroids based on 3D rebuilding environment (3DRE) aims to preserve the biological characteristics of original tumors, but whether the self‐assembly process is influenced by 3DREs remains unknown. We here compared the colorect...

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Main Authors: Yong Zhang, Yang Liu, Yang Yang, Yiwei Li, Lei Liu, Yang Huang, Weiting Zhao, Yong Liu, Minghao Zhang, Yunshan Zhao, Chenggang Li
Format: Article
Language:English
Published: Wiley 2025-07-01
Series:Cancer Medicine
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Online Access:https://doi.org/10.1002/cam4.71017
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author Yong Zhang
Yang Liu
Yang Yang
Yiwei Li
Lei Liu
Yang Huang
Weiting Zhao
Yong Liu
Minghao Zhang
Yunshan Zhao
Chenggang Li
author_facet Yong Zhang
Yang Liu
Yang Yang
Yiwei Li
Lei Liu
Yang Huang
Weiting Zhao
Yong Liu
Minghao Zhang
Yunshan Zhao
Chenggang Li
author_sort Yong Zhang
collection DOAJ
description ABSTRACT Background Three‐dimensional (3D) self‐assembly of organoids or tumoroids based on 3D rebuilding environment (3DRE) aims to preserve the biological characteristics of original tumors, but whether the self‐assembly process is influenced by 3DREs remains unknown. We here compared the colorectal cancer (CRC) tumoroids cultured using different 3DREs, including dome culture (DG), ultra‐low adherence culture without Matrigel(UA) or with Matrigel (UAG), and hanging‐drop without Matrigel (HD) and with Matrigel (HDG). Methods CRC cells were cultured to form tumoroids using DG, UA, UAG, HD, and HDG, respectively. The differences between these tumoroids were examined using light observation, histological staining, RNA sequencing, and drug sensitivity testing. Results The CRC cells aggregated with each other and formed larger, converged tumoroids in the UA and the HD compared to the Matrigel. Histochemical examination revealed that the tumoroids maintained the CRC‐specific characteristics of forming lumens and biomarkers, but the number of lumens decreased, and the cell arrangement was in disorder with increasing impetus of 3DREs that promote cell aggregation ranging from DM, UA to HD. RNA sequencing revealed that the tumoroids retained a similar gene expression pattern, but the oncogenes related to metastasis and poor prognosis were upregulated, and development and morphogenesis‐related genes were downregulated when cultured in the UA and HD compared to the Matrigel. However, the drug sensitivity test showed that the tumoroids, regardless of the methods they were derived, maintained similar sensitivity to drugs. Conclusions We demonstrated that CRC tumoroids developed towards disordered self‐assembly in adaptation to the environment without a matrix, but this adaptation did not alter tumor‐specific phenotypes and drug sensitivity.
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spelling doaj-art-dccfd5fe3bdd4d6a8cf55958aee81ef32025-08-20T02:54:53ZengWileyCancer Medicine2045-76342025-07-011413n/an/a10.1002/cam4.71017Adaptive 3D Self‐Assembly of Colorectal Cancer Cells With Unchanged Tumor Phenotype and Drug SensitivityYong Zhang0Yang Liu1Yang Yang2Yiwei Li3Lei Liu4Yang Huang5Weiting Zhao6Yong Liu7Minghao Zhang8Yunshan Zhao9Chenggang Li10Senior Department of General Surgery, the First Medical Center of Chinese PLA General Hospital Beijing ChinaSenior Department of Hepato‐Pancreato‐Biliary Surgery, the First Medical Center of PLA General Hospital Beijing ChinaDarkjade Sciences, Inc. Beijing ChinaDarkjade Sciences, Inc. Beijing ChinaDarkjade Sciences, Inc. Beijing ChinaDarkjade Sciences, Inc. Beijing ChinaDarkjade Sciences, Inc. Beijing ChinaBeijing Fengtai Hospital of Integrated Traditional Chinese and Western Medicine Beijing ChinaDarkjade Sciences, Inc. Beijing ChinaDarkjade Sciences, Inc. Beijing ChinaSenior Department of General Surgery, the First Medical Center of Chinese PLA General Hospital Beijing ChinaABSTRACT Background Three‐dimensional (3D) self‐assembly of organoids or tumoroids based on 3D rebuilding environment (3DRE) aims to preserve the biological characteristics of original tumors, but whether the self‐assembly process is influenced by 3DREs remains unknown. We here compared the colorectal cancer (CRC) tumoroids cultured using different 3DREs, including dome culture (DG), ultra‐low adherence culture without Matrigel(UA) or with Matrigel (UAG), and hanging‐drop without Matrigel (HD) and with Matrigel (HDG). Methods CRC cells were cultured to form tumoroids using DG, UA, UAG, HD, and HDG, respectively. The differences between these tumoroids were examined using light observation, histological staining, RNA sequencing, and drug sensitivity testing. Results The CRC cells aggregated with each other and formed larger, converged tumoroids in the UA and the HD compared to the Matrigel. Histochemical examination revealed that the tumoroids maintained the CRC‐specific characteristics of forming lumens and biomarkers, but the number of lumens decreased, and the cell arrangement was in disorder with increasing impetus of 3DREs that promote cell aggregation ranging from DM, UA to HD. RNA sequencing revealed that the tumoroids retained a similar gene expression pattern, but the oncogenes related to metastasis and poor prognosis were upregulated, and development and morphogenesis‐related genes were downregulated when cultured in the UA and HD compared to the Matrigel. However, the drug sensitivity test showed that the tumoroids, regardless of the methods they were derived, maintained similar sensitivity to drugs. Conclusions We demonstrated that CRC tumoroids developed towards disordered self‐assembly in adaptation to the environment without a matrix, but this adaptation did not alter tumor‐specific phenotypes and drug sensitivity.https://doi.org/10.1002/cam4.71017adaptive remodelingdrug sensitivityself‐assembly
spellingShingle Yong Zhang
Yang Liu
Yang Yang
Yiwei Li
Lei Liu
Yang Huang
Weiting Zhao
Yong Liu
Minghao Zhang
Yunshan Zhao
Chenggang Li
Adaptive 3D Self‐Assembly of Colorectal Cancer Cells With Unchanged Tumor Phenotype and Drug Sensitivity
Cancer Medicine
adaptive remodeling
drug sensitivity
self‐assembly
title Adaptive 3D Self‐Assembly of Colorectal Cancer Cells With Unchanged Tumor Phenotype and Drug Sensitivity
title_full Adaptive 3D Self‐Assembly of Colorectal Cancer Cells With Unchanged Tumor Phenotype and Drug Sensitivity
title_fullStr Adaptive 3D Self‐Assembly of Colorectal Cancer Cells With Unchanged Tumor Phenotype and Drug Sensitivity
title_full_unstemmed Adaptive 3D Self‐Assembly of Colorectal Cancer Cells With Unchanged Tumor Phenotype and Drug Sensitivity
title_short Adaptive 3D Self‐Assembly of Colorectal Cancer Cells With Unchanged Tumor Phenotype and Drug Sensitivity
title_sort adaptive 3d self assembly of colorectal cancer cells with unchanged tumor phenotype and drug sensitivity
topic adaptive remodeling
drug sensitivity
self‐assembly
url https://doi.org/10.1002/cam4.71017
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