title: Microtubule-targeted drugs inhibit VEGF receptor-2 expression by both transcriptional and post-transcriptional mechanisms.
abstract: Cytoskeletal polymers control a wide range of cellular functions, including proliferation, migration, and gene expression. As changes in endothelial cell shape and motility are required to form vascular networks, we hypothesized that disassembly of actin filaments or microtubules may impact endothelial vascular endothelial growth factor (VEGF) receptor-2 (VEGFR2) expression as a critical determinant of angiogenesis. We therefore investigated the effect of actin filament- and microtubule-disrupting agents on VEGFR1 and VEGFR2 expression by endothelial cells. Microtubule (MT) disassembly greatly inhibited endothelial VEGFR2 expression, whereas VEGFR1 expression levels remained largely unchanged. These suppressive effects were neither conveyed by increased VEGFR2 shedding nor by shortened protein half-life, suggesting that transcriptional mechanisms account for the observed effects. In line with this conclusion, MT disruption significantly suppressed endothelial VEGFR2 mRNA accumulation. The treatment considerably decreased transcriptional activity of 5'-deletional VEGFR2 promoter gene constructs. MT disruption-mediated repression was conveyed by a GC-rich region harboring two consensus Sp1-binding sites. Electrophoretic mobility-shift assay analysis demonstrated that constitutive Sp1-dependent DNA binding is decreased by MT disassembly. In addition, we provide evidence for additional post-transcriptional regulatory mechanisms, as the VEGFR2 mRNA half-life is significantly reduced by MT-disrupting agents. Hence, both inhibition of the rate of gene transcription and increased mRNA turnover represent critical molecular mechanisms by which MT disruption inhibits VEGFR2 expression.
authors.affiliation: Department of Dermatology, Klinikum der Johann Wolfgang Goethe-Universität, Frankfurt am Main, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Meissner
authors.forename: Andreas
authors.initials: A
authors.lastname: Pinter
authors.forename: Despina
authors.initials: D
authors.lastname: Michailidou
authors.forename: Igor
authors.initials: I
authors.lastname: Hrgovic
authors.forename: Natalia
authors.initials: N
authors.lastname: Kaprolat
authors.forename: Monika
authors.initials: M
authors.lastname: Stein
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Holtmeier
authors.forename: Roland
authors.initials: R
authors.lastname: Kaufmann
authors.forename: Jens
authors.initials: J
authors.lastname: Gille
mesh_terms: D000199:Actins / Q000187:drug effects; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002478:Cells, Cultured; D003078:Colchicine / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008870:Microtubules / Q000187:drug effects*; D015739:Nocodazole / Q000494:pharmacology*; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000378:metabolism; D053778:Thiazolidines / Q000494:pharmacology; D014158:Transcription, Genetic; D050257:Tubulin Modulators / Q000494:pharmacology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000187:drug effects / Q000378:metabolism*; D014747:Vinblastine / Q000494:pharmacology*
chemical_list: D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D012333:RNA, Messenger; D016329:Sp1 Transcription Factor; D053778:Thiazolidines; D050257:Tubulin Modulators; D014747:Vinblastine; D040301:Vascular Endothelial Growth Factor Receptor-2; D015739:Nocodazole; D003078:Colchicine; C037067:latrunculin A