Three-dimensional (3D) tumor spheroid invasion assay

J Vis Exp. 2015 May 1;(99):e52686. doi: 10.3791/52686.

Abstract

Invasion of surrounding normal tissues is generally considered to be a key hallmark of malignant (as opposed to benign) tumors. For some cancers in particular (e.g., brain tumors such as glioblastoma multiforme and squamous cell carcinoma of the head and neck - SCCHN) it is a cause of severe morbidity and can be life-threatening even in the absence of distant metastases. In addition, cancers which have relapsed following treatment unfortunately often present with a more aggressive phenotype. Therefore, there is an opportunity to target the process of invasion to provide novel therapies that could be complementary to standard anti-proliferative agents. Until now, this strategy has been hampered by the lack of robust, reproducible assays suitable for a detailed analysis of invasion and for drug screening. Here we provide a simple micro-plate method (based on uniform, self-assembling 3D tumor spheroids) which has great potential for such studies. We exemplify the assay platform using a human glioblastoma cell line and also an SCCHN model where the development of resistance against targeted epidermal growth factor receptor (EGFR) inhibitors is associated with enhanced matrix-invasive potential. We also provide two alternative methods of semi-automated quantification: one using an imaging cytometer and a second which simply requires standard microscopy and image capture with digital image analysis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Brain Neoplasms / pathology*
  • Carcinoma, Squamous Cell / pathology*
  • Cell Culture Techniques / methods*
  • Cell Line, Tumor
  • Glioblastoma / pathology*
  • Head and Neck Neoplasms / pathology*
  • Humans
  • Mice
  • Neoplasm Invasiveness
  • Spheroids, Cellular / pathology
  • Squamous Cell Carcinoma of Head and Neck