title: Selection of suitable reference genes from bone cells in large gradient high magnetic field based on GeNorm algorithm.
abstract: Studies of animals and humans subjected to spaceflight demonstrate that weightlessness negatively affects the mass and mechanical properties of bone tissue. Bone cells could sense and respond to the gravity unloading, and genes sensitive to gravity change were considered to play a critical role in the mechanotransduction of bone cells. To evaluate the fold-change of gene expression, appropriate reference genes should be identified because there is no housekeeping gene having stable expression in all experimental conditions. Consequently, expression stability of ten candidate housekeeping genes were examined in osteoblast-like MC3T3-E1, osteocyte-like MLO-Y4, and preosteoclast-like FLG29.1 cells under different apparent gravities (μg, 1 g, and 2 g) in the high-intensity gradient magnetic field produced by a superconducting magnet. The results showed that the relative expression of these ten candidate housekeeping genes was different in different bone cells; Moreover, the most suitable reference genes of the same cells in altered gravity conditions were also different from that in strong magnetic field. It demonstrated the importance of selecting suitable reference genes in experimental set-ups. Furthermore, it provides an alternative choice to the traditionally accepted housekeeping genes used so far about studies of gravitational biology and magneto biology.
authors: Key Laboratory for Space Bioscience and Biotechnology, Institute of Special Environmental Biophysics, School of Life Science, Northwestern Polytechnical University, Xi'an, China.
authors: Shengmeng
authors: S
authors: Di
authors: Zongcheng
authors: Z
authors: Tian
authors: Airong
authors: A
authors: Qian
authors: Xiang
authors: X
authors: Gao
authors: Dan
authors: D
authors: Yu
authors: Maria Luisa
authors: ML
authors: Brandi
authors: Peng
authors: P
authors: Shang
mesh_terms: D016475:3T3 Cells; D000465:Algorithms; D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods*; D060526:Magnetic Fields; D040542:Mechanotransduction, Cellular / Q000528:radiation effects; D051379:Mice; D010011:Osteocytes / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*