Identification of proteins involved in inhibition of spheroid formation under microgravity.
Many types of cells transit in vitro from a two- to a three-dimensional growth, when they are exposed to microgravity. The underlying mechanisms are not yet understood. Hence, we investigated the impact of microgravity on protein content and growth behavior. For this purpose, the human thyroid cancer cells FTC-133 were seeded either in recently developed cell containers that can endure enhanced physical forces and perform media changes and cell harvesting automatically or in T-25 culture flasks. All cells were cultured for five days at 1g. Afterwards, a part of the cell containers were flown to the International Space Station, while another part was kept on the ground. T-25 flasks were mounted on and next to a Random Positioning Machine. The cells were cultured for 12 days under the various conditions, before they were fixed with RNAlater. All fixed cultures showed monolayers, but three-dimensional aggregates were not detected. In a subsequent protein analysis, 180 proteins were identified by mass spectrometry. These proteins did not indicate significant differences between cells exposed to microgravity and their 1g controls. However, they suggest that an enhanced production of proteins related to the extracellular matrix could detain the cells from spheroid formation, while profilin-1 is phosphorylated.
Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University-Magdeburg, Magdeburg, Germany.
Stefan
S
Riwaldt
Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University-Magdeburg, Magdeburg, Germany.
Jessica
J
Pietsch
Leibniz-Institut für Analytische Wissenschaften -ISAS- e.V, Dortmund, Germany.
Albert
A
Sickmann
Max-Planck Institute for Biochemistry, Martinsried, Germany.
Johann
J
Bauer
Institute for Molecular Physiology and Biotechnology of Plants (IMBIO), Gravitational Biology Group, University of Bonn, Bonn, Germany.
Markus
M
Braun
Airbus, Defense and Space (ADS), Immenstaad, Germany.
Juergen
J
Segerer
Airbus, Defense and Space (ADS), Immenstaad, Germany.
Achim
A
Schwarzwälder
Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University-Magdeburg, Magdeburg, Germany.
Ganna
G
Aleshcheva
Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Thomas J
TJ
Corydon
Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University-Magdeburg, Magdeburg, Germany.
Manfred
M
Infanger
Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Daniela
D
Grimm
D045744:Cell Line, Tumor / Q000378:metabolism / Q000473:pathology; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D009363:Neoplasm Proteins / Q000032:analysis / Q000378:metabolism; D011506:Proteins / Q000032:analysis / Q000378:metabolism*; D018874:Spheroids, Cellular / Q000378:metabolism / Q000473:pathology*; D013964:Thyroid Neoplasms / Q000378:metabolism / Q000473:pathology*; D014893:Weightlessness
D009363:Neoplasm Proteins; D011506:Proteins
Cell biology; Microgravity; Profilin-1; Random Positioning Machine; Spaceflight; Thyroid cancer cells