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Related Experiment Video

Updated: Aug 14, 2025

Time-Resolved Fluorescence Imaging and Analysis of Cancer Cell Invasion in the 3D Spheroid Model
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Time-Resolved Fluorescence Imaging and Analysis of Cancer Cell Invasion in the 3D Spheroid Model

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Visualizing and Quantifying mRNA Localization at the Invasive Front of 3D Cancer Spheroids.

Konstadinos Moissoglu1, Stephen J Lockett2, Stavroula Mili3

  • 1Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|January 18, 2023
PubMed
Summary

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This summary is machine-generated.

Researchers developed a new method to visualize messenger RNA (mRNA) in invading cancer cells. This technique helps understand how mRNA localization drives cell migration in 3D environments.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Cancer Research

Background:

  • Messenger RNA (mRNA) localization at the leading edge of migrating cells is crucial for efficient cell movement.
  • This phenomenon is observed in various contexts, including single cells, 3D multicellular structures, and in vivo tissues.
  • Three-dimensional (3D) cultures offer insights into how extracellular matrix topology and cell-cell interactions affect subcellular mRNA distribution.

Purpose of the Study:

  • To describe a novel method for mRNA imaging within an inducible system of collective cancer cell invasion.
  • To analyze and quantify mRNA distribution at the front of invasive leader cells in a 3D cancer model.

Main Methods:

  • Utilizing MDA-MB-231 cancer cell spheroids embedded in Matrigel.
  • Inducing collective invasion and processing for single-molecule mRNA imaging.
Keywords:
3D invasionCancer spheroidsFluorescence in situ hybridizationFluorescence microscopyFront-back polarityLeader cellMatrigelRNA localizationRNA quantificationSingle-molecule RNA imaging

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  • Employing an analysis algorithm to quantify and compare mRNA distributions.

    • Main Results:

    • Successfully imaged mRNA with single-molecule sensitivity in a 3D collective cancer invasion model.
    • Quantified and compared mRNA distribution at the invasive front of leader cells.
    • Demonstrated the utility of the method in a complex multicellular environment.

    Conclusions:

    • The developed method enables detailed analysis of mRNA localization during collective cell invasion in 3D.
    • This technique can be adapted to study RNA dynamics in other cellular polarization and migration scenarios.
    • Understanding mRNA distribution is key to deciphering mechanisms of cell movement and invasion.