Related Experiment Video
Updated: Oct 3, 2025
Establishing 3-Dimensional Spheroids from Patient-Derived Tumor Samples and Evaluating their Sensitivity to Drugs
Published on: December 16, 2022
Spheroid Culture Differentially Affects Cancer Cell Sensitivity to Drugs in Melanoma and RCC Models
Aleksandra Filipiak-Duliban1,2, Klaudia Brodaczewska1, Arkadiusz Kajdasz3,4
1Laboratory of Molecular Oncology and Innovative Therapies, Military Institute of Medicine, 04-141 Warsaw, Poland.
Abstract:
2D culture as a model for drug testing often turns to be clinically futile. Therefore, 3D cultures (3Ds) show potential to better model responses to drugs observed in vivo. In preliminary studies, using melanoma (B16F10) and renal (RenCa) cancer, we confirmed that 3Ds better mimics the tumor microenvironment. Here, we evaluated how the proposed 3D mode of culture affects tumor cell susceptibility to anti-cancer drugs, which have distinct mechanisms of action (everolimus, doxorubicin, cisplatin). Melanoma spheroids showed higher resistance to all used drugs, as compared to 2D. In an RCC model, such modulation was only observed for doxorubicin treatment. As drug distribution was not affected by the 3D shape, we assessed the expression of MDR1 and mTor. Upregulation of MDR1 in RCC spheroids was observed, in contrast to melanoma. In both models, mTor expression was not affected by the 3D cultures. By NGS, 10 genes related with metabolism of xenobiotics by cytochrome p450 were deregulated in renal cancer spheroids; 9 of them were later confirmed in the melanoma model. The differences between 3D models and classical 2D cultures point to the potential to uncover new non-canonical mechanisms to explain drug resistance set by the tumor in its microenvironment.
Insights
Three-dimensional (3D) cancer cultures better mimic tumor microenvironments than 2D cultures, revealing distinct drug resistance mechanisms. These 3D models show potential for improved preclinical drug testing and understanding of drug resistance.
Area of Science:
- Oncology
- Biomedical Engineering
- Pharmacology
Background:
- Traditional 2D cell cultures often fail to accurately predict in vivo drug responses.
- Three-dimensional (3D) cultures offer a more physiologically relevant model by mimicking the tumor microenvironment.
- Preliminary studies confirmed 3D cultures better represent tumor microenvironments in melanoma and renal cancer models.
Purpose of the Study:
- To evaluate the impact of 3D culture models on tumor cell susceptibility to distinct anti-cancer drugs.
- To investigate the mechanisms underlying altered drug responses in 3D versus 2D cancer models.
- To identify potential non-canonical pathways involved in drug resistance within the tumor microenvironment.
Main Methods:
- Utilized melanoma (B16F10) and renal cell carcinoma (RenCa) cell lines cultured in both 2D and 3D formats.
- Assessed tumor cell susceptibility to anti-cancer drugs: everolimus, doxorubicin, and cisplatin.
- Analyzed drug distribution, MDR1 and mTor expression, and utilized next-generation sequencing (NGS) to identify deregulated genes.
Main Results:
- Melanoma spheroids exhibited increased resistance to all tested drugs compared to 2D cultures.
- Renal cancer spheroids showed modulated resistance only to doxorubicin.
- Upregulation of MDR1 was observed in renal cancer spheroids but not melanoma; mTor expression remained unaffected in both models.
- NGS identified deregulation of 10 xenobiotic metabolism genes (cytochrome P450) in renal cancer spheroids, with 9 confirmed in melanoma spheroids.
Conclusions:
- 3D culture models provide a more accurate representation of tumor response to anti-cancer drugs than 2D models.
- Differences in drug response and gene expression between 2D and 3D cultures highlight the role of the tumor microenvironment in drug resistance.
- 3D models offer a promising platform for uncovering novel mechanisms of drug resistance and improving preclinical drug evaluation.
