title: Loss of miR-200 family in 5-fluorouracil resistant colon cancer drives lymphendothelial invasiveness in vitro.
abstract: Invasive colorectal cancer is associated with poor prognosis requiring treatment with systemic chemotherapies usually including 5-fluorouracil. A consequence of prolonged treatment is the acquisition of resistance eventually resulting in the recurrence of highly metastatic cancer cells. To address the relationship between drug resistance and increased lymphatic metastatic potential, we used a 3D co-culture model of colon tumour cell spheroids of parent CCL227 cells and subclones with gradually increasing resistance against 5-fluorouracil. From each investigated cell line, homogeneous tumour spheroids were generated in the presence of methylcellulose yielding emboli of ∼700 µm diameter. When invasive, tumour spheroids disrupt the continuous lymphendothelial cell (LEC) layer and generate a 'circular chemorepellent-induced defect' (CCID), reminiscent of the entry gates through which tumour emboli intravasate lymphatic vasculature. Here we provide evidence that increasingly chemoresistant colon cancer spheroids were strongly associated with enhanced intravasative properties. In naïve CCL227 spheroids, miR-200 family members were released into exosomes thereby repressing the epithelial to mesenchymal transition-regulating transcription factors ZEB1 and SLUG in LEC. As a consequence of attenuated plasticity and migration of LEC, CCID formation was impaired. Loss of exosomal transferred miR-200c in resistant colon cells rendered LEC more susceptible to pro-migratory signals that were generated and directly transmitted by colon cancer spheroids. This observation indicates a common molecular axis in colon cancer and LEC where miR-200 family members act as regulators of ZEB proteins. The data support the notion that horizontal miR-200 signalling prevents the permeation of cells into adjacent epithelia and contributes to organ integrity.
authors.affiliation: Institute of Clinical Pathology and.
authors.forename: Daniel
authors.initials: D
authors.lastname: Senfter
authors.affiliation: Institute of Clinical Pathology and.
authors.forename: Silvio
authors.initials: S
authors.lastname: Holzner
authors.affiliation: Department of Medicine I, Comprehensive Cancer Center of the Medical University of Vienna, 1090 Vienna, Austria.
authors.forename: Maria
authors.initials: M
authors.lastname: Kalipciyan
authors.affiliation: Department of Medicine I, Comprehensive Cancer Center of the Medical University of Vienna, 1090 Vienna, Austria.
authors.forename: Anna
authors.initials: A
authors.lastname: Staribacher
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, 1090 Vienna, Austria and.
authors.forename: Angelika
authors.initials: A
authors.lastname: Walzl
authors.affiliation: Institute of Clinical Pathology and.
authors.forename: Nicole
authors.initials: N
authors.lastname: Huttary
authors.affiliation: Institute of Clinical Pathology and.
authors.forename: Sigurd
authors.initials: S
authors.lastname: Krieger
authors.affiliation: Department of Clinical Pharmacy and Diagnostics, University of Vienna, 1090 Vienna, Austria.
authors.forename: Stefan
authors.initials: S
authors.lastname: Brenner
authors.affiliation: Department of Clinical Pharmacy and Diagnostics, University of Vienna, 1090 Vienna, Austria.
authors.forename: Walter
authors.initials: W
authors.lastname: Jäger
authors.affiliation: Institute of Clinical Pathology and.
authors.forename: Georg
authors.initials: G
authors.lastname: Krupitza
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, 1090 Vienna, Austria and.
authors.forename: Helmut
authors.initials: H
authors.lastname: Dolznig
authors.affiliation: Department of Medicine I, Comprehensive Cancer Center of the Medical University of Vienna, 1090 Vienna, Austria, robert.mader@meduniwien.ac.at.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Mader
mesh_terms: D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D019008:Drug Resistance, Neoplasm; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D005472:Fluorouracil / Q000494:pharmacology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008207:Lymphatic Metastasis; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D005810:Multigene Family; D009361:Neoplasm Invasiveness; D000071250:Snail Family Transcription Factors; D018874:Spheroids, Cellular / Q000187:drug effects / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured; D000071799:Zinc Finger E-box-Binding Homeobox 1
chemical_list: D018398:Homeodomain Proteins; C533895:MIRN141 microRNA, human; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; C000604407:SNAI1 protein, human; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors; C486307:ZEB1 protein, human; D000071799:Zinc Finger E-box-Binding Homeobox 1; D005472:Fluorouracil