Spheroid Culture Differentially Affects Cancer Cell Sensitivity to Drugs in Melanoma and RCC Models.

Aleksandra Filipiak-Duliban,Klaudia Brodaczewska,Arkadiusz Kajdasz,Claudine Kieda

doi:10.3390/ijms23031166

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.

Highlights

One of the limiting factors to achieve cures for cancer and other diseases is drug resistance
By comparing the 3D cultures with in vivo obtained tumors, we showed that spheroids better mimic the in vivo tumor characteristics than 2D cultures (“Spheroid culture models imitating the tumor microenvironment of renal and melanoma cancer”, submitted)
We showed that our proposed 3D cultures better mimic the in vivo tumor characteristic (“Spheroid culture models imitating the tumor microenvironment of renal and melanoma cancer”, submitted)

Summary

Introduction

One of the limiting factors to achieve cures for cancer and other diseases is drug resistance. This phenomenon is well known, universal, and its development is almost inevitable [1,2]. There are multiple mechanisms underlying cancer unresponsiveness and/or resistance to treatment, reviewed elsewhere [4] They include changes in the drug target (receptor or pathway expression or mutation) [5], induction of metabolism or efflux of drugs [6], interruption of cell death and activation of survival mechanisms [7], and reprogramming the tumor microenvironment to promote tumor growth [8]. A key role played by MDR is the increased activity of drug efflux pumps. This is typically the effect of the ATP-binding cassette (ABCB1/MDR1) [3,9]

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