title: Pathways Regulating Spheroid Formation of Human Follicular Thyroid Cancer Cells under Simulated Microgravity Conditions: A Genetic Approach.
abstract: Microgravity induces three-dimensional (3D) growth in numerous cell types. Despite substantial efforts to clarify the underlying mechanisms for spheroid formation, the precise molecular pathways are still not known. The principal aim of this paper is to compare static 1g-control cells with spheroid forming (MCS) and spheroid non-forming (AD) thyroid cancer cells cultured in the same flask under simulated microgravity conditions. We investigated the morphology and gene expression patterns in human follicular thyroid cancer cells (UCLA RO82-W-1 cell line) after a 24 h-exposure on the Random Positioning Machine (RPM) and focused on 3D growth signaling processes. After 24 h, spheroid formation was observed in RPM-cultures together with alterations in the F-actin cytoskeleton. qPCR indicated more changes in gene expression in MCS than in AD cells. Of the 24 genes analyzed VEGFA, VEGFD, MSN, and MMP3 were upregulated in MCS compared to 1g-controls, whereas ACTB, ACTA2, KRT8, TUBB, EZR, RDX, PRKCA, CAV1, MMP9, PAI1, CTGF, MCP1 were downregulated. A pathway analysis revealed that the upregulated genes code for proteins, which promote 3D growth (angiogenesis) and prevent excessive accumulation of extracellular proteins, while genes coding for structural proteins are downregulated. Pathways regulating the strength/rigidity of cytoskeletal proteins, the amount of extracellular proteins, and 3D growth may be involved in MCS formation.
authors.affiliation: Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke University Clinic, Leipziger Str. 44, 39120 Magdeburg, Germany. stefan.riwaldt@med.ovgu.de.
authors.forename: Stefan
authors.initials: S
authors.lastname: Riwaldt
authors.affiliation: Max Planck Institute for Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany. jbauer@biochem.mpg.de.
authors.forename: Johann
authors.initials: J
authors.lastname: Bauer
authors.affiliation: Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke University Clinic, Leipziger Str. 44, 39120 Magdeburg, Germany. markus.wehland@med.ovgu.de.
authors.forename: Markus
authors.initials: M
authors.lastname: Wehland
authors.affiliation: Institute of Biomedicine, Aarhus University, Wilhelm Meyers Allé 4, 8000 Aarhus C, Denmark. slumstrup@biomed.au.dk.
authors.forename: Lasse
authors.initials: L
authors.lastname: Slumstrup
authors.affiliation: Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke University Clinic, Leipziger Str. 44, 39120 Magdeburg, Germany. sascha.kopp@med.ovgu.de.
authors.forename: Sascha
authors.initials: S
authors.lastname: Kopp
authors.affiliation: Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke University Clinic, Leipziger Str. 44, 39120 Magdeburg, Germany. elisabeth.warnke@med.ovgu.de.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Warnke
authors.affiliation: Institute of Biomedicine, Aarhus University, Wilhelm Meyers Allé 4, 8000 Aarhus C, Denmark. adit88@gmail.com.
authors.forename: Anita
authors.initials: A
authors.lastname: Dittrich
authors.affiliation: Medical Research Laboratory, Department of Clinical Medicine, Faculty of Health, Aarhus University, 8000 Aarhus C, Denmark. nm@clin.au.dk.
authors.forename: Nils E
authors.initials: NE
authors.lastname: Magnusson
authors.affiliation: Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke University Clinic, Leipziger Str. 44, 39120 Magdeburg, Germany. jessica.pietsch@med.ovgu.de.
authors.forename: Jessica
authors.initials: J
authors.lastname: Pietsch
authors.affiliation: Institute of Biomedicine, Aarhus University, Wilhelm Meyers Allé 4, 8000 Aarhus C, Denmark. corydon@biomed.au.dk.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Corydon
authors.affiliation: Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke University Clinic, Leipziger Str. 44, 39120 Magdeburg, Germany. manfred.infanger@med.ovgu.de.
authors.forename: Manfred
authors.initials: M
authors.lastname: Infanger
authors.affiliation: Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke University Clinic, Leipziger Str. 44, 39120 Magdeburg, Germany. dgg@biomed.au.dk.|Institute of Biomedicine, Aarhus University, Wilhelm Meyers Allé 4, 8000 Aarhus C, Denmark. dgg@biomed.au.dk.
authors.forename: Daniela
authors.initials: D
authors.lastname: Grimm
mesh_terms: D018263:Adenocarcinoma, Follicular / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D053263:Gene Regulatory Networks; D006801:Humans; D015398:Signal Transduction; D018874:Spheroids, Cellular; D013961:Thyroid Gland / Q000378:metabolism / Q000473:pathology*; D013964:Thyroid Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D014407:Tumor Cells, Cultured; D018474:Weightlessness Simulation
keywords: caveolin-1; growth; matrix metalloproteinases; pathway studio; random positioning machine; simulated microgravity; thyroid cancer; vascular endothelial growth factor