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Updated: Jan 1, 2026
Therapy Testing in a Spheroid-based 3D Cell Culture Model for Head and Neck Squamous Cell Carcinoma
Published on: April 20, 2018
Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids
Mohammad Azharuddin1, Karin Roberg2,3, Ashis Kumar Dhara4
1Department of Clinical and Experimental Medicine (IKE), Linköping University, Linköping, Sweden.
Abstract:
One of the hallmarks of cancers is their ability to develop resistance against therapeutic agents. Therefore, developing effective in vitro strategies to identify drug resistance remains of paramount importance for successful treatment. One of the ways cancer cells achieve drug resistance is through the expression of efflux pumps that actively pump drugs out of the cells. To date, several studies have investigated the potential of using 3-dimensional (3D) multicellular tumor spheroids (MCSs) to assess drug resistance; however, a unified system that uses MCSs to differentiate between multi drug resistance (MDR) and non-MDR cells does not yet exist. In the present report we describe MCSs obtained from post-diagnosed, pre-treated patient-derived (PTPD) cell lines from head and neck squamous cancer cells (HNSCC) that often develop resistance to therapy. We employed an integrated approach combining response to clinical drugs and screening cytotoxicity, monitoring real-time drug uptake, and assessing transporter activity using flow cytometry in the presence and absence of their respective specific inhibitors. The report shows a comparative response to MDR, drug efflux capability and reactive oxygen species (ROS) activity to assess the resistance profile of PTPD MCSs and two-dimensional (2D) monolayer cultures of the same set of cell lines. We show that MCSs provide a robust and reliable in vitro model to evaluate clinical relevance. Our proposed strategy can also be clinically applicable for profiling drug resistance in cancers with unknown resistance profiles, which consequently can indicate benefit from downstream therapy.
Insights
This study introduces a novel 3D multicellular tumor spheroid model using patient-derived head and neck cancer cells to effectively identify multidrug resistance (MDR). This advanced in vitro system accurately predicts clinical drug resistance in cancer cells.
Area of Science:
- Oncology
- Cancer Biology
- Drug Resistance Research
Background:
- Cancer cells frequently develop resistance to therapeutic agents, necessitating effective in vitro methods for early detection.
- Multidrug resistance (MDR) in cancer is often mediated by efflux pumps that expel therapeutic drugs.
- Existing 3D multicellular tumor spheroid (MCSs) models lack a unified system to differentiate between MDR and non-MDR cells.
Purpose of the Study:
- To develop and validate a 3D multicellular tumor spheroid (MCS) model using patient-derived head and neck squamous cell carcinoma (HNSCC) cell lines.
- To establish a comprehensive strategy for assessing multidrug resistance (MDR) and drug efflux capabilities in cancer cells.
- To compare the drug resistance profiles of 3D MCSs with traditional 2D monolayer cultures.
Main Methods:
- Utilized post-diagnosed, pre-treated patient-derived (PTPD) cell lines from HNSCC.
- Integrated drug response screening, real-time drug uptake monitoring, and transporter activity assessment via flow cytometry.
- Assessed reactive oxygen species (ROS) activity and drug efflux in both 3D MCSs and 2D monolayer cultures.
Main Results:
- The 3D MCS model demonstrated a robust and reliable in vitro system for evaluating drug resistance.
- Comparative analysis revealed distinct resistance profiles between MDR and non-MDR cells within the MCS model.
- The study successfully characterized drug efflux capability and ROS activity in relation to drug resistance.
Conclusions:
- The developed 3D MCS model provides a clinically relevant platform for profiling drug resistance in HNSCC.
- This strategy can aid in identifying patients who may benefit from specific therapeutic interventions.
- The approach offers potential for profiling drug resistance in cancers with previously unknown resistance mechanisms.
