Brain metastases occur in 20 to 40% of patients with metastatic breast cancer. The process is complex and depends on successful cancer cell evasion from the primary tumor, distribution and survival within the blood stream and cerebral microvasculature, penetration of the blood brain barrier and proliferation within the brain microenvironment. The initial steps of brain colonization are difficult to study in vivo. Therefore, in vitro assays have been developed to mimic this process. Most commonly, in vitro studies of brain colonization focus on tumor cell adhesion to brain endothelial cells and transendothelial migration. We previously investigated breast cancer brain colonization from the blood stream in vivo and defined the time and process of brain entry for five different cancer cell lines in a mouse model. We now investigated if in vitro approaches can reliably emulate the initial steps that determine successful brain colonization in vivo. To this end, we optimized an in vitro model of the vascular blood brain barrier and compared the brain invasion properties of the in vivo characterized cell models with their ability to interact with and penetrate the blood brain barrier model in vitro. Our results show that the in vitro findings correlate only poorly with the vivo results. The limitations of the in vitro approaches are discussed in light of the in vivo processes. We conclude that investigation of mechanisms supporting the earliest steps of breast cancer brain metastasis from the blood stream will depend on in vivo analyses.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Al-Shamy G, Sawaya R (2009) Management of brain metastases: the indispensable role of surgery. J Neurooncol 92(3):275–282. doi:10.1007/s11060-009-9839-y
Santarelli JG, Sarkissian V, Hou LC, Veeravagu A, Tse V (2007) Molecular events of brain metastasis. Neurosurg Focus 22(3):E1
Grewal J, Kesari S (2008) Breast cancer surface receptors predict risk for developing brain metastasis and subsequent prognosis. Breast Cancer Res 10(2):104
Grewal J, Zhou H, Factor R, Kesari S (2010) Isolated loss of hormonal receptors in leptomeningeal metastasis from estrogen receptor- and progesterone receptor-positive lobular breast cancer. J Clin Oncol 28(13):200–202. doi:10.1200/JCO.2009.25.3518
DeAngelis LM (2008) Treatment of brain metastasis. J Support Oncol 6(2):87–88
Gerrard GE, Franks KN (2004) Overview of the diagnosis and management of brain, spine, and meningeal metastases. J Neurol Neurosurg Psychiatry 75(Suppl2):ii37–ii42
Norden AD, Wen PY, Kesari S (2005) Brain metastases. Curr Opin Neurol 18(6):654–661
Ballinger WE Jr, Schimpff RD (1979) An experimental model for cerebral metastasis: preliminary light and ultrastructural studies. J Neuropathol Exp Neurol 38(1):19–34
Kawaguchi T, Tobai S, Nakamura K (1982) Extravascular migration of tumor cells in the brain: an electron microscopic study. Invasion Metastasis 2(1):40–50
Paku S, Dome B, Toth R, Timar J (2000) Organ-specificity of the extravasation process: an ultrastructural study. Clin Exp Metastasis 18(6):481–492
Lorger M, Felding-Habermann B (2010) Capturing changes in the brain microenvironment during initial steps of breast cancer brain metastasis. Am J Pathol 176(6):2958–2971. doi:10.2353/ajpath.2010.090838
Kieda C (2003) How endothelial cell organo-specificity mediates circulating cell homing. Arch Immunol Ther Exp (Warsz) 51(2):81–89
Nicolson GL, Menter DG, Herrmann J, Cavanaugh P, Jia L, Hamada J, Yun Z, Nakajima M, Marchetti D (1994) Tumor metastasis to brain: role of endothelial cells, neurotrophins, and paracrine growth factors. Crit Rev Oncog 5(5):451–471
Pauli BU, Lee CL (1988) Organ preference of metastasis. The role of organ-specifically modulated endothelial cells. Lab Invest 58(4):379–387
Palmieri D, Chambers AF, Felding-Habermann B, Huang S, Steeg PS (2007) The biology of metastasis to a sanctuary site. Clin Cancer Res 13(6):1656–1662. doi:10.1158/1078-0432.CCR-06-2659
Palmieri D, Smith QR, Lockman PR, Bronder J, Gril B, Chambers AF, Weil RJ, Steeg PS (2006) Brain metastases of breast cancer. Breast Dis 26:139–147
Bechmann I, Galea I, Perry VH (2007) What is the blood-brain barrier (not)? Trends Immunol 28(1):5–11. doi:10.1016/j.it.2006.11.007
Lee SW, Kim WJ, Park JA, Choi YK, Kwon YW, Kim KW (2006) Blood-brain barrier interfaces and brain tumors. Arch Pharm Res 29(4):265–275
Reichel A, Begley DJ, Abbott NJ (2003) An overview of in vitro techniques for blood-brain barrier studies. Methods Mol Med 89:307–324. doi:10.1385/1-59259-419-0:307
Haseloff RF, Blasig IE, Bauer HC, Bauer H (2005) In search of the astrocytic factor(s) modulating blood-brain barrier functions in brain capillary endothelial cells in vitro. Cell Mol Neurobiol 25(1):25–39
Ribeiro MM, Castanho MA, Serrano I (2010) In vitro blood-brain barrier models–latest advances and therapeutic applications in a chronological perspective. Mini Rev Med Chem 10(3):262–270
Deli MA, Abraham CS, Kataoka Y, Niwa M (2005) Permeability studies on in vitro blood-brain barrier models: physiology, pathology, and pharmacology. Cell Mol Neurobiol 25(1):59–127
Yoneda T, Williams PJ, Hiraga T, Niewolna M, Nishimura R (2001) A bone-seeking clone exhibits different biological properties from the MDA-MB-231 parental human breast cancer cells and a brain-seeking clone in vivo and in vitro. J Bone Miner Res 16(8):1486–1495
Fulcher CA, Zimmerman TS (1982) Characterization of the human factor VIII procoagulant protein with a heterologous precipitating antibody. Proc Natl Acad Sci USA 79(5):1648–1652
Zimmerman TS, Ruggeri ZM, Fulcher CA (1983) Factor VIII/von Willebrand factor. Prog Hematol 13:279–309
Bellail AC, Hunter SB, Brat DJ, Tan C, Van Meir EG (2004) Microregional extracellular matrix heterogeneity in brain modulates glioma cell invasion. Int J Biochem Cell Biol 36(6):1046–1069. doi:10.1016/j.biocel.2004.01.013
Giese A, Westphal M (1996) Glioma invasion in the central nervous system. Neurosurgery 39(2):235–250 discussion 250–232
Gaillard PJ, Voorwinden LH, Nielsen JL, Ivanov A, Atsumi R, Engman H, Ringbom C, de Boer AG, Breimer DD (2001) Establishment and functional characterization of an in vitro model of the blood-brain barrier, comprising a co-culture of brain capillary endothelial cells and astrocytes. Eur J Pharm Sci 12(3):215–222
Rubin LL, Hall DE, Porter S, Barbu K, Cannon C, Horner HC, Janatpour M, Liaw CW, Manning K, Morales J et al (1991) A cell culture model of the blood-brain barrier. J Cell Biol 115(6):1725–1735
Kis B, Deli MA, Kobayashi H, Abraham CS, Yanagita T, Kaiya H, Isse T, Nishi R, Gotoh S, Kangawa K, Wada A, Greenwood J, Niwa M, Yamashita H, Ueta Y (2001) Adrenomedullin regulates blood-brain barrier functions in vitro. Neuroreport 12(18):4139–4142
Schulze C, Smales C, Rubin LL, Staddon JM (1997) Lysophosphatidic acid increases tight junction permeability in cultured brain endothelial cells. J Neurochem 68(3):991–1000
Carbonell WS, Ansorge O, Sibson N, Muschel R (2009) The vascular basement membrane as “soil” in brain metastasis. PLoS One 4(6):e5857. doi:10.1371/journal.pone.0005857
Bergers G, Brekken R, McMahon G, TH Vu, Itoh T, Tamaki K, Tanzawa K, Thorpe P, Itohara S, Werb Z, Hanahan D (2000) Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. Nat Cell Biol 2(10):737–744. doi:10.1038/35036374
Bos PD, Zhang XH, Nadal C, Shu W, Gomis RR, Nguyen DX, Minn AJ, van de Vijver MJ, Gerald WL, Foekens JA, Massague J (2009) Genes that mediate breast cancer metastasis to the brain. Nature 459(7249):1005–1009. doi:10.1038/nature08021
Supported by grants from the National Institute of Health (CA112287 to BFH), Department of Defense (W81XWH-08-1-0468 to BFH), California Breast Cancer Research Program (15IB-0050, 16IB-0052 to BFH and 16IB-0030 to ML), and a Susan G. Komen Breast Cancer Foundation fellowship (PDF0707935 to ML).
Electronic supplementary material
Below is the link to the electronic supplementary material.
About this article
Cite this article
Lorger, M., Lee, H., Forsyth, J.S. et al. Comparison of in vitro and in vivo approaches to studying brain colonization by breast cancer cells. J Neurooncol 104, 689–696 (2011). https://doi.org/10.1007/s11060-011-0550-4
- Brain metastasis
- Breast cancer
- In vitro
- In vivo
- Blood brain barrier
- Transendothelial migration
- Brain metastasis model