title: Identifications of novel mechanisms in breast cancer cells involving duct-like multicellular spheroid formation after exposure to the Random Positioning Machine.
abstract: Many cell types form three-dimensional aggregates (MCS; multicellular spheroids), when they are cultured under microgravity. MCS often resemble the organ, from which the cells have been derived. In this study we investigated human MCF-7 breast cancer cells after a 2 h-, 4 h-, 16 h-, 24 h- and 5d-exposure to a Random Positioning Machine (RPM) simulating microgravity. At 24 h few small compact MCS were detectable, whereas after 5d many MCS were floating in the supernatant above the cells, remaining adherently (AD). The MCS resembled the ducts formed in vivo by human epithelial breast cells. In order to clarify the underlying mechanisms, we harvested MCS and AD cells separately from each RPM-culture and measured the expression of 29 selected genes with a known involvement in MCS formation. qPCR analyses indicated that cytoskeletal genes were unaltered in short-term samples. IL8, VEGFA, and FLT1 were upregulated in 2 h/4 h AD-cultures. The ACTB, TUBB, EZR, RDX, FN1, VEGFA, FLK1 Casp9, Casp3, PRKCA mRNAs were downregulated in 5d-MCS-samples. ESR1 was upregulated in AD, and PGR1 in both phenotypes after 5d. A pathway analysis revealed that the corresponding gene products are involved in organization and regulation of the cell shape, in cell tip formation and membrane to membrane docking.
authors.affiliation: Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von Guericke-University, D-39120 Magdeburg, Germany.
authors.forename: Sascha
authors.initials: S
authors.lastname: Kopp
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Lasse
authors.initials: L
authors.lastname: Slumstrup
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Corydon
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Jayashree
authors.initials: J
authors.lastname: Sahana
authors.affiliation: Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von Guericke-University, D-39120 Magdeburg, Germany.
authors.forename: Ganna
authors.initials: G
authors.lastname: Aleshcheva
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Tawhidul
authors.initials: T
authors.lastname: Islam
authors.affiliation: Medical Research Laboratory, Department of Clinical Medicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Nils E
authors.initials: NE
authors.lastname: Magnusson
authors.affiliation: Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von Guericke-University, D-39120 Magdeburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Wehland
authors.affiliation: Max-Planck-Institute of Biochemistry, D-82152 Martinsried, Germany.
authors.forename: Johann
authors.initials: J
authors.lastname: Bauer
authors.affiliation: Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von Guericke-University, D-39120 Magdeburg, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Infanger
authors.affiliation: Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von Guericke-University, D-39120 Magdeburg, Germany.|Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Daniela
authors.initials: D
authors.lastname: Grimm
mesh_terms: D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D061986:MCF-7 Cells; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D025941:Protein Interaction Mapping; D015398:Signal Transduction; D018874:Spheroids, Cellular / Q000378:metabolism* / Q000648:ultrastructure; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics / Q000378:metabolism; D018474:Weightlessness Simulation / Q000295:instrumentation*
chemical_list: C508606:CXCL8 protein, human; D003598:Cytoskeletal Proteins; C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha; D016209:Interleukin-8; D047908:Intracellular Signaling Peptides and Proteins; C439067:MRFAP1 protein, human; D009687:Nuclear Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C501162:FLT1 protein, human; D040281:Vascular Endothelial Growth Factor Receptor-1