Abstract
The methods described here provide a standardized process for assessing in vitro tumor cell migration and invasion in real time. The kinetic data generated under these standardized conditions are reproducible and characteristic of individual tumor cell lines. The complex kinetic features of the data can be analyzed using parameters modeled after pharmacokinetic data processing. Application of the method to the array of tumor types included in the National Cancer Institute’s sixty cell line panel (NCI60) revealed distinct modes of invasion with some tumor cell lines utilizing a mesenchymal mode and generating information-rich kinetic profiles. Other cell lines utilized an amoeboid mode not suitable for detection with this method. The method described will be useful as a guide for tumor cell line selection and as a starting point in designing experiments probing migration and invasion.
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Acknowledgments
The authors thank Adam S. Harned, Kunio Nagashima and Ulrich Baxa for expert assistance with scanning electron microscopy and John Connelly for help with siRNA experiments. Dr. Dominic Scudiero, Anne Monks, Eric Harris and Thomas E. Silvers provided technical and data processing support. This project has been funded in whole or in part with federal funds from the National Cancer Institute, National Institutes of Health, under Contract HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. government. This work was supported in part by Roche Applied Science.
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1 Electronic Supplementary Material
Supplementary Fig. 1
Individual phenotypic data for all of the NCI60 tumor cell lines. Graphic data are presented as an interactive MS Excel spreadsheet. Individual tumor cell lines can be selected in the box at the upper right and the corresponding data will display. This MS Excel file, with an embedded macro, has been tested on PCs running Windows XP. It does not work on Macintosh computers. (XLS 4970 kb)
Supplementary Table 1
Listing of the average invasion area under the curve (AUC) for the NCI60 tumor cell lines. Cell line names are given, as well as tumor panel assignment, associated panel numbers, and cell numbers. (XLS 28 kb)
Supplementary Table 2
Correlation of in vitro invasion phenotype, based on area under the curve (AUC), with molecular characteristics of the NCI60 tumor cell lines. Associations are presented in rank order, with the highest correlation at the top. Some portions of the characterization database remain confidential, thus the top listed correlation is 3 and there are skips in the listing. Most column headings are self-explanatory. PCC = Pearson correlation coefficient. (XLS 53 kb)
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DeLosh, R.M., Shoemaker, R.H. (2021). Evaluation of Real-Time In Vitro Invasive Phenotypes. In: Stein, U.S. (eds) Metastasis. Methods in Molecular Biology, vol 2294. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1350-4_12
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DOI: https://doi.org/10.1007/978-1-0716-1350-4_12
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