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

  1. Author:
    Moissoglu, Konstadinos
    Lockett,Stephen
    Mili, Stavroula

  2. Author Address

    Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, USA., Optical Microscopy and Analysis Laboratory, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research Inc. for the National Cancer Institute, NIH, Frederick, MD, USA., Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, USA. voula.mili@nih.gov.,
    1. Year: 2023
  2. Journal: Methods in Molecular Biology (Clifton, N.J.)
    1. 2608
    2. Pages: 263-280
  4. Type of Article: Article
  1. Abstract:

    Localization of mRNAs at the front of migrating cells is a widely used mechanism that functionally supports efficient cell movement. It is observed in single cells on two-dimensional surfaces, as well as in multicellular three-dimensional (3D) structures and in tissue in vivo. 3D multicellular cultures can reveal how the topology of the extracellular matrix and cell-cell contacts influence subcellular mRNA distributions. Here we describe a method for mRNA imaging in an inducible system of collective cancer cell invasion. MDA-MB-231 cancer cell spheroids are embedded in Matrigel, induced to invade, and processed to image mRNAs with single-molecule sensitivity. An analysis algorithm is used to quantify and compare mRNA distributions at the front of invasive leader cells. The approach can be easily adapted and applied to analyze RNA distributions in additional settings where cells polarize along a linear axis. © 2023. The Authors.

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  2. DOI: 10.1007/978-1-0716-2887-4_16
  3. PMID: 36653713

Library Notes

  1. Fiscal Year: FY2022-2023
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