title: Cancer-associated fibroblast promote transmigration through endothelial brain cells in three-dimensional in vitro models.
abstract: Brain metastases are associated with high morbidity as well as with poor prognosis and survival in breast cancer patients. Despite its clinical importance, metastasis of breast cancer cells through the blood-brain barrier (BBB) is poorly understood. The objective of our study was to investigate whether cancer-associated fibroblasts (CAFs) play crucial roles in breast cancer brain metastasis. Using a cell adhesion assays, in vitro BBB permeability and transmigration assays and soft agar colony formation assays, we investigated the physical roles of CAFs in breast cancer brain metastasis. We also performed immunofluorescence, flow cytometric analysis, Droplet Digital PCR and Simon™ Simple Western System to confirm changes in expression levels. We established two novel three-dimensional (3D) culture systems using a perpendicular slide chamber and applying 3D embedded culture method to reflect brain metastasis conditions. With a newly developed device, CAFs was proven to promote cell adhesion to human brain microvascular endothelial cells, in vitro BBB permeability and transmigration and colony formation of breast cancer cells. Furthermore, CAFs enhanced the invasive migration of breast cancer cells in two kinds of 3D cultures. These 3D models also reliably recapitulate the initial steps of BBB transmigration, micro-metastasis and colonization. Expression of integrin α5β1 and αvβ3, c-MET and α2,6-siayltransferase was increased in breast cancer cells that migrated through the BBB. In conclusion, based on our in vitro BBB and co-culture models, our data suggest that CAFs may play a role in breast cancer brain metastasis.
authors.affiliation: Department of Chemistry, National Creative Research Center for Biofunctional Molecules, Yonsei University, Seoul, Korea; Department of Chemistry, Brain Korea 21 PLUS Project, Yonsei University, Seoul, Korea.
authors.forename: Yoon Pyo
authors.initials: YP
authors.lastname: Choi
authors.forename: Joo Hyun
authors.initials: JH
authors.lastname: Lee
authors.forename: Ming-Qing
authors.initials: MQ
authors.lastname: Gao
authors.forename: Baek Gil
authors.initials: BG
authors.lastname: Kim
authors.forename: Suki
authors.initials: S
authors.lastname: Kang
authors.forename: Se Hoon
authors.initials: SH
authors.lastname: Kim
authors.forename: Nam Hoon
authors.initials: NH
authors.lastname: Cho
mesh_terms: D001692:Biological Transport; D001812:Blood-Brain Barrier; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism / Q000473:pathology*; D001932:Brain Neoplasms / Q000235:genetics / Q000378:metabolism / Q000556:secondary*; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002448:Cell Adhesion; D002465:Cell Movement; D049109:Cell Proliferation; D018920:Coculture Techniques; D004730:Endothelium, Vascular / Q000378:metabolism / Q000473:pathology*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000378:metabolism / Q000473:pathology*; D014407:Tumor Cells, Cultured
chemical_list: D012333:RNA, Messenger
keywords: 3D culture; blood-brain barrier; brain metastasis; breast cancer; cancer-associated fibroblast