Selection of suitable reference genes from bone cells in large gradient high magnetic field based on GeNorm algorithm
Selection of suitable reference genes from bone cells in large gradient high magnetic field based on GeNorm algorithm
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DOI：
10.3109/15368378.2011.608869
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发表时间：
2011-01-01
影响因子：
1.7
通讯作者：
Shang, Peng
中科院分区：
文献类型：
--
作者：
Di, Shengmeng;Tian, Zongcheng;Shang, Peng
对遭受太空飞行的动物和人类的研究表明，失重对骨组织的质量和机械性能产生负面影响。骨细胞能够感知和响应重力卸载，对重力变化敏感的基因被认为在骨细胞的机械转导中起着关键作用。由于在所有实验条件下都没有稳定表达的内参基因，因此要评估基因表达的折叠变化，需要确定合适的内参基因。因此，在超导磁体产生的高强度梯度磁场中，在不同的表观重力（mu g， 1g和2g）下，研究了10个候选看家基因在成骨细胞样MC3T3-E1、骨细胞样MLO-Y4和破骨细胞前样FLG29.1细胞中的表达稳定性。结果表明，这10个候选管家基因在不同骨细胞中的相对表达量不同；此外，同种细胞在变重力条件下的最适内参基因与在强磁场条件下的最适内参基因也不同。这表明在实验设置中选择合适的内参基因的重要性。此外，它为传统上公认的内务基因提供了另一种选择，目前用于引力生物学和磁生物学的研究。
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 (mu g, 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.