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Comparison of EMT-Related and Multi-Drug Resistant Gene Expression, Extracellular Matrix Production, and Drug
Xiaoli Qi1,2, Alexandra V Prokhorova1, Alexander V Mezentsev1
1School of Biological and Medical Physics, Moscow Institute of Physics and Technology, National Research University, 141701 Dolgoprudny, Russia.
Abstract:
Multicellular 3D tumor models are becoming a powerful tool for testing of novel drug products and personalized anticancer therapy. Tumor spheroids, a commonly used 3D multicellular tumor model, more closely reproduce the tumor microenvironment than conventional 2D cell cultures. It should be noted that spheroids can be produced using different techniques, which can be subdivided into scaffold-free (SF) and scaffold-based (SB) methods. However, it remains unclear, to what extent spheroid properties depend on the method of their generation. In this study, we aimed to carry out a head-to-head comparison of drug sensitivity and molecular expression profile in SF and SB spheroids along with a monolayer (2D) cell culture. Here, we produced non-small cell lung cancer (NSCLC) spheroids based on human lung adenocarcinoma cell line A549. Drug sensitivity analysis of the tested cell cultures to five different chemotherapeutics resulted in IC50 (A549-SB) > IC50 (A549-SF) > IC50 (A549-2D) trend. It was found that SF and SB A549 spheroids displayed elevated expression levels of epithelial-to-mesenchymal transition (EMT) markers and proteins associated with drug resistance compared with the monolayer A549 cell culture. Enhanced drug resistance of A549-SB spheroids can be a result of larger diameters and elevated deposition of extracellular matrix (ECM) that impairs drug penetration into spheroids. Thus, the choice of the spheroid production method can influence the properties of the generated 3D cell culture and their drug resistance. This fact should be considered for correct interpretation of drug testing results.
Insights
Scaffold-free and scaffold-based 3D tumor spheroids show increased drug resistance compared to 2D cultures. Spheroid generation method impacts drug sensitivity and molecular profiles, crucial for accurate drug testing.
Area of Science:
- Oncology
- 3D Cell Culture Technology
- Drug Discovery and Development
Background:
- Multicellular 3D tumor models, such as tumor spheroids, offer a more relevant tumor microenvironment compared to 2D cell cultures.
- Tumor spheroids can be generated using scaffold-free (SF) or scaffold-based (SB) methods, but their impact on spheroid properties is not fully understood.
Purpose of the Study:
- To conduct a head-to-head comparison of drug sensitivity and molecular expression profiles between SF and SB spheroids and 2D cell cultures.
- To investigate the influence of spheroid generation methods on drug resistance in non-small cell lung cancer (NSCLC) models.
Main Methods:
- Generation of non-small cell lung cancer (NSCLC) spheroids using the human lung adenocarcinoma cell line A549 via scaffold-free and scaffold-based methods.
- Drug sensitivity analysis using five different chemotherapeutics to determine IC50 values for 2D cultures, SF spheroids, and SB spheroids.
- Analysis of molecular expression profiles, focusing on epithelial-to-mesenchymal transition (EMT) markers and drug resistance proteins.
Main Results:
- A trend of increasing drug resistance was observed: IC50 (A549-SB) > IC50 (A549-SF) > IC50 (A549-2D).
- Both SF and SB A549 spheroids exhibited elevated expression of EMT markers and drug resistance proteins compared to 2D cultures.
- Enhanced drug resistance in A549-SB spheroids may be attributed to larger diameters and increased extracellular matrix deposition, hindering drug penetration.
Conclusions:
- The method used for spheroid generation significantly influences the properties and drug resistance of 3D cell cultures.
- Scaffold-based spheroids demonstrated higher drug resistance, potentially due to structural factors affecting drug penetration.
- Consideration of the spheroid production method is essential for the accurate interpretation of drug testing results in 3D tumor models.
