Common Effects on Cancer Cells Exerted by a Random Positioning Machine and a 2D Clinostat |
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Authors: | Benjamin Svejgaard Markus Wehland Xiao Ma Sascha Kopp Jayashree Sahana Elisabeth Warnke Ganna Aleshcheva Ruth Hemmersbach Jens Hauslage Jirka Grosse Johann Bauer Thomas Juhl Corydon Tawhidul Islam Manfred Infanger Daniela Grimm |
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Institution: | 1. Department of Biomedicine, Aarhus University, Aarhus, Denmark.; 2. Clinic for Plastic, Aesthetic and Hand Surgery, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.; 3. DLR, German Aerospace Center, Institute of Aerospace Medicine, Cologne, Germany.; 4. Department of Nuclear Medicine, University of Regensburg, Regensburg, Germany.; 5. Max-Planck Institute for Biochemistry, Martinsried, Germany.; Osaka University Graduate School of Medicine, JAPAN, |
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Abstract: | 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. |
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