Loss of cancer drug activity in colon cancer HCT-116 cells during spheroid formation in a new 3-D spheroid cell culture system
Abstract
Clinically relevant in vitro methods are needed to identify new cancer drugs for solid tumors. We report on a new 3-D spheroid cell culture system aimed to mimic the properties of solid tumors in vivo. The colon cancer cell lines HCT-116 wt and HCT-116 wt/GFP were grown as monolayers and for 3 or 6 days on 96-well NanoCulture® plates to form spheroids. Expression of surface markers, genes and hypoxia were assessed to characterize the spheroids and drug induced cytotoxicity was evaluated based on fluorescein diacetate (FDA) conversion by viable cells to fluorescent fluorescein or by direct measurement of fluorescence of GFP marked cells after a 72 h drug incubation. The cells reproducibly formed spheroids in the NanoCulture® plates with tight cell-attachment after 6 days. Cells in spheroids showed geno- and phenotypical properties reminiscent of hypoxic stem cells. Monolayer cultured cells were sensitive to standard and investigational drugs, whereas the spheroids gradually turned resistant. Similar results for cytotoxicity were observed using simplified direct measurement of fluorescence of GFP marked cells compared with FDA incubation. In conclusion, this new 3-D spheroid cell culture system provides a convenient and clinically relevant model for the identification and characterization of cancer drugs for solid tumors.
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In HCT-116 colon cancer cells, all four standard anticancer drugs and two investigational drugs were highly active in 2D monolayer culture but generally less active and gradually lost activity in 3D spheroids, with 6-day spheroids more resistant than 3-day spheroids.
“The results indicated that all drugs were highly active in 2D monolayer culture but generally less active and gradually lost their activity in 3D spheroids (6 day spheroids were more resistant than 3 day spheroids in Fig. 4 ), irrespective of different action mechanisms”
From Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors (2014) · cites this paper as [56]
Cells cultured in 3D models are more resistant to anticancer drugs than 2D cultures.
“A number of studies have found that cells cultured in 3D models are more resistant to anticancer drugs than 2D cultures. 41,56”
From Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors (2014) · cites this paper as [56]
The results in the 3D model more closely reflected in vivo observations.
“The results in the 3D model more closely reflected in vivo observations.”
From In Vitro research Reproducibility: Keeping up High Standards (2019) · cites this paper as Karlsson et al., 2012
HCT-116 cells in 3D culture displayed increased resistance against anti-cancer drugs compared with the 2D model.
“These aspects were exemplified by a study using HCT-116 cells in 3D culture that displayed increased resistance against anti-cancer drugs compared with the 2D model (Karlsson et al., 2012).”
From In Vitro research Reproducibility: Keeping up High Standards (2019) · cites this paper as Karlsson et al., 2012
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References 39 · Cited by 180
- Authors
- Karlsson, H., Fryknäs, M., Larsson, R., & Nygren, P.
- Affiliations
- Uppsala University (SE)
- Corresponding author
- Karlsson, Henning
- Venue
- Experimental Cell Research
- Publisher
- Elsevier BV
- Published
- 2012-08
- Volume
- 318
- Issue
- 13
- Pages
- 1577-1585
- ISSN
- 0014-4827 (ISSN-L)
- OpenAlex
- W2075889966
- Cited by
- 180
- References
- 39
- Citations by year
- 2026: 2 · 2025: 16 · 2024: 11 · 2023: 13 · 2022: 17 · 2021: 15 · 2020: 11 · 2019: 17 · 2018: 19 · 2017: 19 · 2016: 11 · 2015: 9 · 2014: 8 · 2013: 10 · 2012: 2
- FWCI
- 4.59
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Experimental Cell Research (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Cancerfonden
- Topics
- Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Keywords
- colon cancer, HCT116 cells, drug resistance, drug sensitivity, hypoxia, cancer stem cells, solid tumor models, GFP-labeled cells, fluorescein diacetate, cytotoxicity assay, tumor microenvironment
- MeSH terms
- Antineoplastic Agents, Cell Survival, Colonic Neoplasms, Drug Screening Assays, Antitumor, Humans, Immunohistochemistry, Cell Hypoxia, Gene Expression, Spheroids, Cellular, Cell Culture Techniques, Drug Resistance, Neoplasm, HCT116 Cells, Green Fluorescent Proteins
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