3D Spheroid Model: A Smarter Way to Study Treatment Resistance in Head and Neck Cancer
Posted on 2026-04-27
Head and Neck Squamous Cell Carcinoma (HNSCC) is the 7th most common cancer globally, with up to 870,000 new cases and 440,000 deaths every year [1]. Unlike many other cancers, HNSCC tumours can develop in diverse areas within the head and neck- including the oral and nasal cavities, pharynx and larynx [2,3]. This diversity makes HNSCC one of the most complex malignancies, with different patients responding differently to the same treatment. This article will introduce the concept of using 3D spheroid culture model as a tool to study these treatment responses and its potential in guiding more personalised therapies.
Figure 1: Anatomical regions where HNSCC can develop. Tumours typically originate from the mucosal epithelium within the head and neck.
Over the recent decade, the use of 3D spheroid model in research has gained increasing popularity over traditional 2D culture. One key advantage comes to its improved physiological relevance. The 3D model has the ability to reproduce complex tumour architectures, such as oxygen gradient, cell-cell interaction, and drug diffusion patterns [4], which would otherwise be challenging to capture using conventional 2D culture. The 3D system also offers the flexibility to model interactions with the tumour microenvironment, such as extracellular matrix and immune system cross-talks, both of which can have strong influence over a tumour’s response to drug treatments [5]. In preclinical research, the 3D spheroid model acts as a valuable bridge between the flat 2D culture and sophisticated animal models - generating a system that is more biologically accurate than the former, yet more scalable and ethically accessible than the latter.
Figure 2: Key advantages of the use of 3D spheroid model in the modern cancer research landscape, compared to conventional 2D culture and animal model.
An exciting application of using a 3D spheroid model in HNSCC research is in studying molecular mechanisms of drug resistance. Resistance to chemotherapy occurs in up to 33% of HNSCC patients, and is the major contributor to treatment failure and poor survival outcomes [6]. Using this model, drug-resistant cell populations can first be generated in a conventional 2D culture system through prolonged drug exposure, typically for a period ranging from 6 to 9 months. These resistant populations can then be used to generate 3D spheroid model, often displaying an altered morphology and growth pattern from their drug-sensitive counterpart. This 3D spheroid model provides powerful visual and quantitative insights into drug response, including visibly increased cell death at higher doses, and elevated drug tolerance confirmed by cell viability assays.
Figure 3: Experimental workflow of generating an in vitro model to study response to chemotherapy drug using a HNSCC cell line. Workflow involves first generating chemo-resistant cell populations (2D), then using these populations to create a 3D, chemo-resistant HNSCC spheroid model. From the point of cell seeding into ultra-low attachment plate, the 3D spheroid takes 72 hours to form. The fully formed 3D spheroids were then dosed with cisplatin (a platinum-based chemotherapy drug) for a further 72 hours, prior to examining its response to drug exposure.
Beyond that, the 3D spheroid model is also compatible with high-throughput downstream analyses to further characterise the complex mechanisms driving treatment resistant in HNSCC. These include RNA Sequencing to identify biomarkers associated with drug-resistant subtypes [7], paving its way for targeted therapeutic discovery tailored for each population of patients. Additionally, the high scalability and reproducibility offer an ideal platform for drug screening to identify candidate compounds capable of mitigating treatment resistance.
As cancer research continues to evolve, there is an increasing focus on developing patient-relevant models that can better reflect the biology within human tumours. The 3D spheroid model serves as a major advancement from the conventional 2D culture system, aligning with the 3Rs principle (replacement, reduction, refinement) and the changing regulatory landscape [8]. Hence, the 3D spheroid model is gaining increased recognition in translational research from bench to bedside, paving its way to more ground-breaking discoveries in cancer research.
Acknowledgement
I would like to thank my PhD supervisor, Prof. Muy-Teck Teh, and Queen Mary University of London for their ongoing guidance throughout my project, and for their valuable feedback in shaping this article.
Written by Hailey Sze (PhD Researcher, Queen Mary University of London)
References
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