Skip to main content
SpringerLink
Log in

Evaluation of Real-Time In Vitro Invasive Phenotypes

  • Protocol
  • First Online:
Metastasis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2294))

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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Talmadge JE, Fidler IJ (2010) AACR centennial series: the biology of cancer metastasis: historical perspective. Cancer Res 70(14):5649–5669

    Article  CAS  Google Scholar 

  2. Boyden S (1962) The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes. J Exp Med 115:453–466

    Article  CAS  Google Scholar 

  3. Albini A, Iwamoto Y, Kleinman HK, Martin GR, Aaronson SA, Kozlowski JM (1987) McEwan RN. A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res 47(12):3239–3245

    CAS  PubMed  Google Scholar 

  4. Imamura H, Takao S (1994) Aikou T. A modified invasion-3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide assay for quantitating tumor cell invasion. Cancer Res 54(13):3620–3624

    CAS  PubMed  Google Scholar 

  5. Sieuwerts AM, Klijn JG, Foekens JA (1997) Assessment of the invasive potential of human gynecological tumor cell lines with the in vitro Boyden chamber assay: influences of the ability of cells to migrate through the filter membrane. Clin Exp Metastasis 15(1):53–62

    Article  CAS  Google Scholar 

  6. Solly K, Wang X, Xu X, Strulovici B, Zheng W (2004) Application of real-time cell electronic sensing (RT-CES) technology to cell-based assays. Assay Drug Dev Technol 2(4):363–372

    Article  CAS  Google Scholar 

  7. Xing JZ, Zhu L, Jackson JA, Gabos S, Sun XJ, Wang XB, Xu X (2005) Dynamic monitoring of cytotoxicity on microelectronic sensors. Chem Res Toxicol 18(2):154–161

    Article  CAS  Google Scholar 

  8. Monks A, Scudiero D, Skehan P, Shoemaker R, Paull K, Vistica D, Hose C, Langley J, Cronise P, Vaigro-Wolff A, Gray-Goodrich M, Campbell H, Mayo J, Boyd M (1991) Feasibility of a high-flux anticancer drug screen utilizing a diverse panel of cultured human tumor cell lines. J Natl Cancer Inst 83:757–766

    Article  CAS  Google Scholar 

  9. Paull KD, Shoemaker RH, Hodes L, Monks A, Scudiero DA, Rubinstein L, Plowman J, Boyd MR (1989) Display and analysis of patterns of differential activity of drugs against human tumor cell lines: development of mean graph and COMPARE algorithm. J Natl Cancer Inst 81:1088–1092

    Article  CAS  Google Scholar 

  10. Shoemaker RH (2006) The NCI60 human tumour cell line anticancer drug screen. Nat Rev Cancer 6:813–823

    Article  CAS  Google Scholar 

  11. Friedl P, Wolf K (2010) Plasticity of cell migration: a multiscale tuning model. J Cell Biol 188(1):11–19

    Article  CAS  Google Scholar 

  12. Vajkoczy P, Knyazev P, Kunkel A, Capelle HH, Behrndt S, von Tengg-Kobligk H, Kiessling F, Eichelsbacher U, Essig M, Read TA, Erber R, Ullrich A (2006) Dominant-negative inhibition of the Axl receptor tyrosine kinase suppresses brain tumor cell growth and invasion and prolongs survival. Proc Natl Acad Sci U S A 103(15):5799–5804

    Article  CAS  Google Scholar 

  13. Suyama K, Shapiro I, Guttman M, Hazan RB (2002) A signaling pathway leading to metastasis is controlled by N-cadherin and the FGF receptor. Cancer Cell 2(4):301–314. Mol Cancer Ther. 2008 Oct;7(10):3408-19

    Article  CAS  Google Scholar 

  14. Fischer H, Taylor N, Allerstorfer S, Grusch M, Sonvilla G, Holzmann K, Setinek U, Elbling L, Cantonati H, Grasl-Kraupp B, Gauglhofer C, Marian B, Micksche M, Berger W (2008) Fibroblast growth factor receptor-mediated signals contribute to the malignant phenotype of non-small cell lung cancer cells: therapeutic implications and synergism with epidermal growth factor receptor inhibition. Mol Cancer Ther 7:3408–3419

    Article  CAS  Google Scholar 

  15. Reinhold WC, Sunshine M, Liu H, Varma S, Kohn KW, Morris J, Doroshow J, Pommier Y (2012) CellMiner: a web-based suite of genomic and pharmacologic tools to explore transcript and drug patterns in the NCI-60 cell line set. Cancer Res 72(14):3499–3511

    Article  CAS  Google Scholar 

  16. Sahai E (2005) Mechanisms of cancer cell invasion. Curr Opin Genet Dev 15:87–96

    Article  CAS  Google Scholar 

  17. Wosikowski K, Schuurhuis D, Johnson K, Paull KD, Myers TG, Weinstein JN, Bates SE (1997) Identification of epidermal growth factor receptor and c-erbB2 pathway inhibitors by correlation with gene expression patterns. J Natl Cancer Inst 89:1505–1515

    Article  CAS  Google Scholar 

  18. Yigitbasi OG, Younes MN, Doan D, Jasser SA, Schiff BA, Bucana CD, Bekele BN, Fidler IJ, Myers JN (2004) Tumor cell and endothelial cell therapy of oral cancer by dual tyrosine kinase receptor blockade. Cancer Res 64(21):7977–7984

    Article  CAS  Google Scholar 

  19. Hannigan G, Troussard AA, Dedhar S (2005) Integrin-linked kinase: a cancer therapeutic target unique among its ilk. Nat Rev Cancer 5:51–63

    Article  CAS  Google Scholar 

Download references

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.

Author information

Authors and Affiliations

  1. Leidos Biomedical Research, Frederick National Laboratory for Cancer Research, Frederick, MD, USA

    René M. DeLosh

  2. Division of Cancer Prevention, National Cancer Institute, Bethesda, MD, USA

    Robert H. Shoemaker

Authors
  1. René M. DeLosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Robert H. Shoemaker
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Robert H. Shoemaker .

Editor information

Editors and Affiliations

  1. Translational Oncology of Solid Tumors Experimental and Clinical Research Center, Charité – Universitätsmedizin Berlin and Max-Delbrück-Center for Molecular Medicine, Berlin, Germany

    Ulrike S. Stein

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)

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

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

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1350-4_12

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1349-8

  • Online ISBN: 978-1-0716-1350-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation