Common Effects on Cancer Cells Exerted by a Random Positioning Machine and a 2D Clinostat.
In this study we focused on gravity-sensitive proteins of two human thyroid cancer cell lines (ML-1; RO82-W-1), which were exposed to a 2D clinostat (CLINO), a random positioning machine (RPM) and to normal 1g-conditions. After a three (3d)- or seven-day-culture (7d) on the two devices, we found both cell types growing three-dimensionally within multicellular spheroids (MCS) and also cells remaining adherent (AD) to the culture flask, while 1g-control cultures only formed adherent monolayers, unless the bottom of the culture dish was covered by agarose. In this case, the cytokines IL-6 and IL-8 facilitated the formation of MCS in both cell lines using the liquid-overlay technique at 1g. ML-1 cells grown on the RPM or the CLINO released amounts of IL-6 and MCP-1 into the supernatant, which were significantly elevated as compared to 1g-controls. Release of IL-4, IL-7, IL-8, IL-17, eotaxin-1 and VEGF increased time-dependently, but was not significantly influenced by the gravity conditions. After 3d on the RPM or the CLINO, an accumulation of F-actin around the cellular membrane was detectable in AD cells of both cell lines. IL-6 and IL-8 stimulation of ML-1 cells for 3d and 7d influenced the protein contents of ß1-integrin, talin-1, Ki-67, and beta-actin dose-dependently in adherent cells. The ß1-integrin content was significantly decreased in AD and MCS samples compared with 1g, while talin-1 was higher expressed in MCS than AD populations. The proliferation marker Ki-67 was elevated in AD samples compared with 1g and MCS samples. The ß-actin content of R082-W-1 cells remained unchanged. ML-1 cells exhibited no change in ß-actin in RPM cultures, but a reduction in CLINO samples. Thus, we concluded that simulated microgravity influences the release of cytokines in follicular thyroid cancer cells, and the production of ß1-integrin and talin-1 and predicts an identical effect under real microgravity conditions.
Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Benjamin
B
Svejgaard
Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.
Markus
M
Wehland
Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Xiao
X
Ma
Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Sascha
S
Kopp
Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Jayashree
J
Sahana
Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.
Elisabeth
E
Warnke
Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.
Ganna
G
Aleshcheva
DLR, German Aerospace Center, Institute of Aerospace Medicine, Cologne, Germany.
Ruth
R
Hemmersbach
DLR, German Aerospace Center, Institute of Aerospace Medicine, Cologne, Germany.
Jens
J
Hauslage
Department of Nuclear Medicine, University of Regensburg, Regensburg, Germany.
Jirka
J
Grosse
Max-Planck Institute for Biochemistry, Martinsried, Germany.
Johann
J
Bauer
Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Thomas Juhl
TJ
Corydon
Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Tawhidul
T
Islam
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
D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D045744:Cell Line, Tumor; D016207:Cytokines / Q000096:biosynthesis*; D006801:Humans; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D018874:Spheroids, Cellular / Q000378:metabolism*; D014893:Weightlessness
D016207:Cytokines; D009363:Neoplasm Proteins