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Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures

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Published on: December 26, 2016

Optimization of OSCC spheroid generation for high-throughput drug screening.

Mansi Dobariya¹, Seena Sunil¹, Sai Swetha Uppalapati¹

¹Department of Biotechnology, National Institute of Pharmaceutical Education and Research, Palaj, Gandhinagar, Gujarat 382355 India.

In Vitro Models

|February 19, 2026

Summary

Three-dimensional (3D) spheroid models using ultra-low attachment (ULA) plates offer a reproducible method for studying oral squamous cell carcinoma (OSCC) biology. These 3D models exhibit increased chemoresistance, highlighting their value for preclinical drug screening.

Keywords:

3D spheroid model Drug screening Oral squamous cell carcinoma (OSCC)Scaffold-free culture Ultra-low attachment (ULA) plates

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Area of Science:

Oncology
Biotechnology
Cancer Research

Background:

Three-dimensional (3D) spheroid models more accurately mimic the in vivo tumor microenvironment compared to traditional two-dimensional (2D) cultures.
This makes 3D spheroids valuable tools for preclinical drug screening in oral squamous cell carcinoma (OSCC).

Purpose of the Study:

To establish and characterize scaffold-free 3D spheroid models for OSCC using different culture methods.
To compare the drug response of 3D spheroids with 2D cultures for OSCC cell lines.

Main Methods:

Scaffold-free spheroids were generated from CAL 27 and OECM-1 OSCC cell lines using methylcellulose suspension, agarose-coated plates, and ultra-low attachment (ULA) plates.
Morphology, uniformity, viability, necrosis, gene expression of cancer hallmark genes, and drug response to cisplatin and doxorubicin were assessed.

Main Results:

Compact, uniform spheroids formed within 72 hours, with ULA plates yielding the highest reproducibility and structural integrity.
3D spheroids exhibited central necrosis, modulated cancer hallmark gene expression (hypoxia, angiogenesis, EMT), and significantly higher resistance to cisplatin and doxorubicin compared to 2D cultures.

Conclusions:

ULA-based 3D spheroids provide a reproducible, stable, and scalable model that accurately reflects OSCC tumor biology and chemoresistance.
These 3D models are suitable for high-throughput drug screening and advancing translational cancer research in OSCC.