Reduced Expression of Cytoskeletal and Extracellular Matrix Genes in Human Adult Retinal Pigment Epithelium Cells Exposed to Simulated Microgravity

被引：54
作者：
Corydon, Thomas J. [1 ]
Mann, Vivek [2 ]
Slumstrup, Lasse [1 ]
Kopp, Sascha [3 ]
Sahana, Jayashree [1 ]
Askou, Anne Louise [1 ]
Magnusson, Nils E. [4 ]
Echegoyen, David [3 ]
Bek, Toke [5 ]
Sundaresan, Alamelu [6 ]
Riwaldt, Stefan [1 ]
Bauer, Johann [6 ]
Infanger, Manfred [3 ]
Grimm, Daniela [1 ,3 ]
机构：
[1] Aarhus Univ, Dept Biomed, Wilhelm Meyers Alle 4, DK-8000 Aarhus C, Denmark
[2] Texas Southern Univ, Dept Biol, Houston, TX 77004 USA
[3] Otto Von Guericke Univ, Clin Plast Aesthet & Hand Surg, Magdeburg, Germany
[4] Aarhus Univ, Dept Clin Med, Med Res Lab, Aarhus, Denmark
[5] Aarhus Univ Hosp, Dept Ophthalmol, Aarhus, Denmark
[6] Max Planck Inst Biochem, Martinsried, Germany
来源：
CELLULAR PHYSIOLOGY AND BIOCHEMISTRY | 2016年 / 40卷 / 1-2期
关键词：
Simulated microgravity; Retinal pigment epithelium cells; 3D growth; Spheroids; Gene expression; Random Positioning Machine; Cytoskeleton; Vascular endothelial growth factor; ENDOTHELIAL GROWTH-FACTOR; THYROID-CANCER CELLS; SPHEROID FORMATION; 3-DIMENSIONAL GROWTH; PARABOLIC FLIGHT; CHOROIDAL FOLDS; GRAVITY; DIFFERENTIATION; CULTURE; MECHANISMS;
D O I：
10.1159/000452520
中图分类号：
Q2 [细胞生物学];
学科分类号：
071009 ; 090102 ;
摘要：
Background/Aims: Microgravity (mu g) has adverse effects on the eye of humans in space. The risk of visual impairment is therefore one of the leading health concerns for NASA. The impact of mu g on human adult retinal epithelium (ARPE-19) cells is unknown. Methods: In this study we investigated the influence of simulated pg (s-mu g; 5 and 10 days (d)), using a Random Positioning Machine (RPM), on ARPE-19 cells. We performed phase-contrast/ fluorescent microscopy, qRT-PCR, Western blotting and pathway analysis. Results: Following RPM-exposure a subset of ARPE-19 cells formed multicellular spheroids (MCS), whereas the majority of the cells remained adherent (AD). After 5d, alterations of F-actin and fibronectin were observed which reverted after 10d-exposure, suggesting a time-dependent adaptation to s-pg. Gene expression analysis of 12 genes involved in cell structure, shape, adhesion, migration, and angiogenesis suggested significant changes after a 10d-RPM-exposure. 11 genes were down-regulated in AD and MCS 10d-RPM-samples compared to lg, whereas FLK1 was up-regulated in 5d- and 10d-RPM-MCS-samples. Similarly, TIMP1 was up-regulated in 5d-RPM-samples, whereas the remaining genes were down-regulated in 5d-RPM-samples. Western blotting revealed similar changes in VEGF, 8-actin, laminin and fibronectin of 5d-RPM-samples compared to 10d, whereas different alterations of p-tubulin and vimentin were observed. The pathway analysis showed complementing effects of VEGF and integrin beta-1. Conclusions: These findings clearly show that s-mu g induces significant alterations in the F-actin-cytoskeleton and cytoskeleton-related proteins of ARPE-19, in addition to changes in cell growth behavior and gene expression patterns involved in cell structure, growth, shape, migration, adhesion and angiogenesis. (C) 2016 The Author(s) Published by S. Karger AG, Basel
引用
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页码：1 / 17
页数：17
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