title: Inhibition of tumour spheroid-induced prometastatic intravasation gates in the lymph endothelial cell barrier by carbamazepine: drug testing in a 3D model.
abstract: Metastatic breast cancer is linked to an undesired prognosis. One early and crucial metastatic step is the interaction of cancer emboli with adjacent stroma or endothelial cells, and understanding the mechanisms of this interaction provides the basis to define new targets as well as drugs for therapy and disease management. A three-dimensional (3D) co-culture model allowing the examination of lymphogenic dissemination of breast cancer cells was recently developed which facilitates not only the study of metastatic processes but also the testing of therapeutic concepts. This 3D setting consists of MCF-7 breast cancer cell spheroids (representing a ductal and hormone-dependent subtype) and of hTERT-immortalised lymph endothelial cell (LEC; derived from foreskin) monolayers. Tumour spheroids repel the continuous LEC layer, thereby generating "circular chemorepellent-induced defects" (CCIDs) that are reminiscent to the entry gates through which tumour emboli intravasate lymphatics. We found that the ion channel blocker carbamazepine (which is clinically used to treat epilepsy, schizophrenia and other neurological disorders) inhibited CCID formation significantly. This effect correlated with the inhibition of the activities of NF-κB, which contributes to cell motility, and with the inactivation of the mobility proteins MLC2, MYPT1 and FAK which are necessary for LEC migration. NF-κB activity and cell movement are prerequisites of CCID formation. On the other hand, the expression of the motility protein paxillin and of the NF-κB-dependent adhesion mediator ICAM-1 was unchanged. Also the activity of ALOX12 was unaffected. ALOX12 is the main enzyme synthesising 12(S)-HETE, which then triggers CCID formation. The relevance of the inhibition of CYP1A1, which is also involved in the generation of mid-chain HETEs such as 12(S)-HETE, by carbamazepine remains to be established, because the constitutive level of 12(S)-HETE did not change upon carbamazepine treatment. Nevertheless, the effect of carbamazepine on the inhibition of CCID formation as an early step of breast cancer metastasis was significant and substantial (~30 %) and achieved at concentrations that are found in the plasma of carbamazepine-treated adults (40-60 μM). The fact that carbamazepine is a drug approved by the US Food and Drug Administration facilitates a "from-bench-to-bedside" perspective. Therefore, the here presented data should undergo scrutiny in vivo.
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Mathias
authors.initials: M
authors.lastname: Teichmann
authors.forename: Nicole
authors.initials: N
authors.lastname: Kretschy
authors.forename: Sabine
authors.initials: S
authors.lastname: Kopf
authors.forename: Kanokwan
authors.initials: K
authors.lastname: Jarukamjorn
authors.forename: Atanas G
authors.initials: AG
authors.lastname: Atanasov
authors.forename: Katharina
authors.initials: K
authors.lastname: Viola
authors.forename: Benedikt
authors.initials: B
authors.lastname: Giessrigl
authors.forename: Philipp
authors.initials: P
authors.lastname: Saiko
authors.forename: Thomas
authors.initials: T
authors.lastname: Szekeres
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Mikulits
authors.forename: Verena M
authors.initials: VM
authors.lastname: Dirsch
authors.forename: Nicole
authors.initials: N
authors.lastname: Huttary
authors.forename: Sigurd
authors.initials: S
authors.lastname: Krieger
authors.forename: Walter
authors.initials: W
authors.lastname: Jäger
authors.forename: Michael
authors.initials: M
authors.lastname: Grusch
authors.forename: Helmut
authors.initials: H
authors.lastname: Dolznig
authors.forename: Georg
authors.initials: G
authors.lastname: Krupitza
mesh_terms: D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / Q000378:metabolism; D001092:Arachidonate 12-Lipoxygenase / Q000378:metabolism; D002220:Carbamazepine / Q000494:pharmacology*; D024722:Cardiac Myosins / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D018920:Coculture Techniques; D019363:Cytochrome P-450 CYP1A1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects*; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism; D006801:Humans; D061986:MCF-7 Cells / Q000187:drug effects / Q000473:pathology; D018994:Myosin Light Chains / Q000378:metabolism; D043263:Myosin-Light-Chain Phosphatase / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D018874:Spheroids, Cellular / Q000187:drug effects
chemical_list: D018994:Myosin Light Chains; D016328:NF-kappa B; C107184:myosin light chain 2; D002220:Carbamazepine; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D001092:Arachidonate 12-Lipoxygenase; C476558:ALOX12 protein, human; C569463:CYP1A1 protein, human; D019363:Cytochrome P-450 CYP1A1; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human; D043263:Myosin-Light-Chain Phosphatase; C490372:PPP1R12A protein, human; D024722:Cardiac Myosins