Methods for Studying Tumor Cell Migration and Invasiveness
Authors:
P. Kovaříková 1; E. Michalová 2; L. Knopfová 3; P. Bouchal 1,2
Authors‘ workplace:
Ústav biochemie, Přírodovědecká fakulta MU, Brno
1; Regionální centrum aplikované molekulární onkologie, Masarykův onkologický ústav, Brno
2; Ústav experimentální biologie, Přírodovědecká fakulta MU, Brno
3
Published in:
Klin Onkol 2014; 27(Supplementum): 22-27
Overview
Migration and invasiveness are phenotypic characteristics of cells that contribute to physiological processes, such as wound healing or embryogenesis and they are involved in serious pathological processes, namely in tumor cell metastasis. Availability of methods for studying migration and invasiveness of the cells is important for understanding molecular basis of these processes. In the case of cancer, migration, invasiveness and metastatic potential of tumor cells are key factors that determine clinical prognosis of the patients. This communication provides an overview of in vitro and in vivo methods which are used to study cell migration, invasion and metastasis. In vitro methods for studying cell migration include simple two‑dimensional assays (scratch – wound assay and the assay based on the effect of hepatocyte growth factor) and methods based on chemotaxis (Dunn‘s chamber, videomicroscopy of cells, the use of carriers with chemoattractants). Methods for studying both cell migration and invasiveness in vitro include more complex systems based on the principle of the Boyden chamber (transwell migration/ invasive test, analysis of cell migration and invasion in xCELLigence system, confocal microscopy based approaches) as well as analysis of cell migration in micro‑channels. Our overview of in vivo methods provides an introduction into model organisms and methods used in this field, with an emphasis on the study of cancer metastasis in mouse models. The methods described in this review are mainly involved in larger research projects aiming at developing new diagnostic and therapeutic approaches in oncology.
Key words:
migration – invasiveness – in vitro assays – in vivo models – metastasis – tumor cells
This work was supported by the project of Czech Science Foundation No. 14-19250S, by the European Regional Development Fund and the State Budget of the Czech Republic (RECAMO, CZ.1.05/2.1.00/03.0101) and by MH CZ – DRO (MMCI, 00209805).
The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.
The Editorial Board declares that the manuscript met the ICMJE “uniform requirements” for biomedical papers.
Submitted:
27. 1. 2014
Accepted:
31. 3. 2014
Sources
1. Vincente‑ Manzanares M, Horwitz AR. Cell migration: an overview. Methods Mol Biol 2011; 769: 1−24. doi: 10.1007/ 978-1-61779- 207- 6_1.
2. Valastyan S, Weinberg RA. Tumor metastasis: molecular insights and evolving paradigms. Cell 2011; 147(2): 275– 292. doi: 10.1016/ j.cell.2011.09.024.
3. Patel LR, Camacho DF, Shiozawa Y et al. Mechanisms of cancer cell metastasis to the bone: a multistep process. Future Oncol 2011; 7(11): 1285– 1297. doi: 10.2217/ fon.11.112.
4. Penet MF, Chen Z, Bhujwalla ZM. MRI of metastasis permissive microenvironments. Future Oncol 2011; 7(11): 1269– 1284. doi: 10.2217/ fon.11.114.
5. Maryáš J, Faktor J, Dvořáková M et al. Proteomics in investigation of cancer metastasis: Functional and clinical consequences and methodological challenges. Proteomics 2014; 14(4−5): 426−440. doi: 10.1002/ pmic.201300264.
6. Faktor J, Dvorakova M, Maryas J et al. Identification and characterization of pro‑metastatic targets, pathways and molecular complexes using a toolbox of proteomic technology. Klin Onkol 2012; 25 (Suppl 2): 2S70−2S77.
7. Hughes CS, Postovit LM, Lajoie GA. Matrigel: a complex protein mixture required for optimal growth of cell culture. Proteomics 2010; 10(9): 1886−1890. doi: 10.1002/ pmic.200900758.
8. Cory G. Scratch‑ wound assay. Methods Mol Biol 2011; 769: 25−30. doi: 10.1007/ 978-1- 61779- 207- 6_2.
9. Magdalena J, Millard TH, Etienne‑ Manneville S et al. Involvement of the Arp2/ 3 Complex and Scar2 in Golgi Polarity in scratch wound models. Mol Biol Cell 2003; 14(2): 670−684.
10. Eccles SA, Box C, Court W. Cell migration/ invasion assays and their application in cancer drug discovery. Biotechnol Annu Rev 2005; 11: 391– 421.
11. Knopfová L. Funkce proteinu c‑ Myb ve vybraných aspektech kancerogeneze. Informační listy GSGM 2013; 41: 37−50.
12. Fram TS, Wells CM, Jones GE. HGF‑induced DU145 cell scatter assay. Methods Mol Biol 2011; 769: 31−40. doi: 10.1007/ 978- 1- 61779- 207- 6_3.
13. Thiery JP. Epithelial‑ mesenchymal transitions in tumour progression. Nat Rev Cancer 2002; 2(6): 442−454.
14. Cooper CR, Pienta KJ. Cell adhesion and chemotaxis in prostate cancer metastasis to bone: a minireview. Prostate Cancer Prostatic Dis 2000; 3(1): 6−12.
15. Zicha D, Dunn GA, Brown AS. A new direct‑ viewing chemotaxis chamber. J Cell Sci 1991; 9(Pt 4): 769−775.
16. Kassis J, Lauffenburger DA, Turner T et al. Tumor invasion as dysregulated cell motility. Semin Cancer Biol 2001; 11(2): 105−117.
17. Zantl R, Horn E. Chemotaxis of slow migrating mammalian cells analysed by video microscopy. Methods Mol Biol 2011; 769: 191−203. doi: 10.1007/ 978- 1- 61779- 207- 6_13.
18. Le Y, Zhou Y, Iribarren P et al. Chemokines and chemokine receptors: their manifold roles in homeostasis and disease. Cell Mol Immunol 2004; 1(2): 95−104.
19. Theveneau E, Mayor R. Beads on the run: beads as alternative tools for chemotaxis assays. Methods Mol Biol 2011; 769: 449−460. doi: 10.1007/ 978- 1- 61779- 207- 6_30.
20. Falasca M, Raimondi C, Maffucci T. Boyden chamber. Methods Mol Biol 2011; 769: 87−95. doi: 10.1007/ 978- 1- 61779- 207- 6_7.
21. Boyden S. The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leukocytes. J Exp Med 1962; 115: 453−466.
22. Marshall J. Transwell® invasion assays. Methods Mol Biol 2011; 769: 97−110. doi: 10.1007/ 978- 1- 61779- 207- 6_8.
23. Limame R, Wouters A, Pauwels B et al. Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real‑ time technology and classic endpoint assays. PLoS One 2012; 7(10): e46536. doi: 10.1371/ journal.pone.0046536.
24. Aceabio.com [homepage on the Internet]. CA: Acea Biosciences, Inc.; c2014 [cited 2014 January 25]. Available from: http:/ / www.aceabio.com/ product_info.aspx?id=184.
25. Bird C, Kirstein S. Real‑ time, label‑free monitoring of cellular invasion and migration with the xCELLigence system. Nature Methods 2009; 6: v‑ vi.
26. RTCA DP Instrument Operator’s Manual. Germany: Roche Diagnostics 2009.
27. Cain RJ, Borda d’Água B, Ridley AJ. Quantification of transendothelial migration using three‑ dimensional confocal microscopy. Methods Mol Biol 2011; 769: 167−190. doi: 10.1007/ 978- 1- 61779- 207-6_12.
28. Friedl P, Wolf K. Plasticity of cell migration: a multiscale tunning model. J Cell Biol 2010; 188(1): 11−19.
29. Heuzé ML, Collin O, Terriac E et al. Cell migration in confinement: a micro‑channel‑based assay. Methods Mol Biol 2011; 769: 415−434. doi: 10.1007/ 978- 1- 61779- 207- 6_28.
30. Artemenko Y, Swaney KF, Devreotes PN. Assessment of development and chemotaxis in Dictyostelium discodeum mutants. Methods Mol Biol 2011; 769: 287−309. doi: 10.1007/ 978- 1- 61779- 207- 6_20.
31. Wong M, Martynovsky M, Schwarzbauer JE. Analysis of cell migration using Ceanorhabditis elegans as a model system. Methods Mol Biol 2011; 769: 233−248. doi: 10.1007/ 978- 1- 61779- 207- 6_16.
32. Stramer B, Wood W. Inflammation and wound healing in Drosophila. Methods Mol Biol 2009; 571: 137−149. doi: 10.1007/ 978- 1- 60761- 198- 1_9.
33. Xu T, Rubin TM. Analysis of genetic mosaics in developing and adult Drosophila tissues. Development 1993; 117(4): 1223−1237.
34. Lieschke GJ, Currie PD. Animal models of human disease: zebrafish swim into view. Nat Rev Genet 2007; 8(5): 353−367.
35. Elks PM, Loynes CA, Renshaw SA. Measuring inflammatory cell migration in the zebrafis. Methods Mol Biol 2011; 769: 261−275. doi: 10.1007/ 978- 1- 61779- 207- 6_18.
36. Box GM, Eccles SA. Simple experimental and spontaneous metastatic assays in mice. Methods Mol Biol 2011; 769: 311−329. doi: 10.1007/ 978- 1- 61779-207- 6_21.
37. Lu J, Steeg PS, Price JE et al. Breast cancer metastasis: challenges an opportunities. Cancer Res 2009; 69(12): 4951−4953. doi: 10.1158/ 0008- 5472.CAN‑ 09- 0099.
38. Talmadge JE. Models of metastasis in drug discovery. Methods Mol Biol 2010; 602: 215−233. doi: 10.1007/ 978- 1- 60761- 058- 8_13.
39. Aslakson CJ, Miller FR. Selective events in the metastatic proces defined by analysis of the sequential dissemination of subpopulations of a mouse mammary tumor. Cancer Res 1992; 52(6): 1399−1405.
40. Price JE, Polyzos A, Zhang RD et al. Tumorigenicity and metastasis of human breast carcinoma cell lines in nude mice. Cancer Res 1990; 50(3): 717−721.
41. Havens AM, Pedersen EA, Shiozawa Y et al. An in vivo mouse model for human prostate cancer metastasis. Neoplasia 2008; 10(4): 371−380.
Labels
Paediatric clinical oncology Surgery Clinical oncologyArticle was published in
Clinical Oncology
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