Abstract
For normal embryonic development/morphogenesis, cell migration and homing are well-orchestrated and important events requiring specific cellular mechanisms. In diseases such as cancer deregulated cell migration represents a major problem. Therefore, numerous efforts are under way to understand the molecular mechanisms of tumor cell migration and to generate more efficient tumor therapies. Cell migration assays are one of the most commonly used functional assays. The wound-healing assay or the Boyden chamber assay are variations of these assays. Nearly all of them are two-dimensional assays and the cells can only migrate on one substrate at a time. This is in contrast to the in vivo situation where the cells are faced simultaneously with different surfaces and interact with different cell types. To approach this in vivo situation we used a modified version of the stripe assay designed by Bonhoeffer and colleagues to examine mechanisms of axonal guidance. The design of this assay allows cells to decide between two different substrates offered at the same time. Utilizing alternating neuronal substrates for migration analyses we can partially mimic the complex in vivo situation for brain tumor cells. Here we describe the detailed protocol to perform a modified version of the stripe assay in order to observe substrate-dependent migration effects in vitro, to analyze the effect of Rho-dependent kinases (ROCKS), of histone deacetylases (HDACs) and of other molecules on glioma cells.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kramer N, Walzl A, Unger C, Rosner M, Krupitza G, Hengstschlager M, Dolznig H (2013) In vitro cell migration and invasion assays. Mutat Res 752(1):10–24
Gibbons HM, Dragunow M (2010) Adult human brain cell culture for neuroscience research. Int J Biochem Cell Biol 42(6):844–856
Filipovic R, Kumar SS, Bahr BA, Loturco J (2014) Slice culture method for studying migration of neuronal progenitor cells derived from human embryonic stem cells (hESC). Curr Protoc Stem Cell Biol 29:1H.7.1–1H.7.14
Walter J, Kern-Veits B, Huf J, Stolze B, Bonhoeffer F (1987) Recognition of position-specific properties of tectal cell membranes by retinal axons in vitro. Development 101(4):685–696
Walter J, Henke-Fahle S, Bonhoeffer F (1987) Avoidance of posterior tectal membranes by temporal retinal axons. Development 101(4):909–913
Knoll B, Weinl C, Nordheim A, Bonhoeffer F (2007) Stripe assay to examine axonal guidance and cell migration. Nat Protoc 2(5):1216–1224
Mertsch S, Oellers P, Wendling M, Stracke W, Thanos S (2013) Dissecting the inter-substrate navigation of migrating glioblastoma cells with the stripe assay reveals a causative role of ROCK. Mol Neurobiol 48(1):169–179
Mertsch S, Thanos S (2014) Opposing signaling of ROCK1 and ROCK2 determines the switching of substrate specificity and the mode of migration of glioblastoma cells. Mol Neurobiol 49(2):900–915
Oellers P, Schroer U, Senner V, Paulus W, Thanos S (2009) ROCKs are expressed in brain tumors and are required for glioma-cell migration on myelinated axons. Glia 57(5):499–509
Wang C, Chowdhury S, Driscoll M, Parent CA, Gupta SK, Losert W (2014) The interplay of cell-cell and cell-substrate adhesion in collective cell migration. J R Soc Interface 11(100):20140684
Maxwell GD (1976) Substrate dependence of cell migration from explanted neural tubes in vitro. Cell Tissue Res 172(3):325–330
Shahbazian MD, Grunstein M (2007) Functions of site-specific histone acetylation and deacetylation. Annu Rev Biochem 76:75–100
Koutsounas I, Giaginis C, Theocharis S (2013) Histone deacetylase inhibitors and pancreatic cancer: are there any promising clinical trials? World J Gastroenterol 19(8):1173–1181
Muller S, Kramer OH (2010) Inhibitors of HDACs—effective drugs against cancer? Curr Cancer Drug Targets 10(2):210–228
Huang L (2006) Targeting histone deacetylases for the treatment of cancer and inflammatory diseases. J Cell Physiol 209(3):611–616
Carew JS, Giles FJ, Nawrocki ST (2008) Histone deacetylase inhibitors: mechanisms of cell death and promise in combination cancer therapy. Cancer Lett 269(1):7–17
Watanabe M, Adachi S, Matsubara H, Imai T, Yui Y, Mizushima Y, Hiraumi Y, Watanabe K, Kamitsuji Y, Toyokuni SY, Hosoi H, Sugimoto T, Toguchida J, Nakahata T (2009) Induction of autophagy in malignant rhabdoid tumor cells by the histone deacetylase inhibitor FK228 through AIF translocation. Int J Cancer 124(1):55–67
Zhu L, Wu K, Ma S, Zhang S (2015) HDAC inhibitors: a new radiosensitizer for non-small-cell lung cancer. Tumori 101:257–262
Eigl BJ, North S, Winquist E, Finch D, Wood L, Sridhar SS, Powers J, Good J, Sharma M, Squire JA, Bazov J, Jamaspishvili T, Cox ME, Bradbury PA, Eisenhauer EA, Chi KN (2015) A phase II study of the HDAC inhibitor SB939 in patients with castration resistant prostate cancer: NCIC clinical trials group study IND195. Invest New Drugs 33(4):969–976
Ji M, Lee EJ, Kim KB, Kim Y, Sung R, Lee SJ, Kim DS, Park SM (2015) HDAC inhibitors induce epithelial-mesenchymal transition in colon carcinoma cells. Oncol Rep 33(5):2299–2308
Lin KT, Wang YW, Chen CT, Ho CM, Su WH, Jou YS (2012) HDAC inhibitors augmented cell migration and metastasis through induction of PKCs leading to identification of low toxicity modalities for combination cancer therapy. Clin Cancer Res 18(17):4691–4701
Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, Scheithauer BW, Kleihues P (2007) The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol 114(2):97–109
Messaoudi K, Clavreul A, Lagarce F (2015) Toward an effective strategy in glioblastoma treatment. Part I: resistance mechanisms and strategies to overcome resistance of glioblastoma to temozolomide. Drug Discov Today 20(7):899–905
Acknowledgement
This work was performed in the Institute of Experimental Ophthalmology, Muenster, with the technical assistance of M. Wissing and M. Langkamp-Flock and the support of Dr. M. Wendling.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Mertsch, S., Thanos, S. (2017). Analysis of Histone Deacetylase-Dependent Effects on Cell Migration Using the Stripe Assay. In: Krämer, O. (eds) HDAC/HAT Function Assessment and Inhibitor Development. Methods in Molecular Biology, vol 1510. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6527-4_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6527-4_5
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6525-0
Online ISBN: 978-1-4939-6527-4
eBook Packages: Springer Protocols