南方医科大学学报

南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (04): 516-.

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Importin 8在成骨分化中的细胞内定位及表达差异

郎斌，汪鑫平，车向新，吴萍，许晓源   

  • 出版日期:2013-04-20 发布日期:2013-04-20

  • Online:2013-04-20 Published:2013-04-20

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摘要： 目的观察Importin 8（IPO8）在成骨分化过程中的细胞内定位与表达差异。方法对人成骨样SaOS-2细胞进行成骨诱导
分化，采用茜素红染色、免疫细胞化学方法和实时定量PCR技术检测成骨分化效果及分化过程中IPO8的细胞内定位和表达差
异。结果茜素红染色显示SaOS-2细胞成骨诱导第10天可见大量红色结节样结构。免疫细胞化学方法显示，IPO8主要定位于
细胞核周胞质。诱导至第3天，IPO8胞浆胞核均有表达，但明显开始向核内移动。至第7天，核内表达更明显，而到第10天，细
胞呈骨细胞样，IPO8均匀分布于胞质内。实时定量PCR技术检测发现，OPN基因在SaOS-2细胞成骨分化过程中表达增加，而
IPO8基因在成骨分化至第3天时表达最高，之后呈下降趋势。结论IPO8在成骨分化过程中的细胞内定位与表达存在明显差
异，提示IPO8可能参与成骨细胞分化过程的调控。

Abstract: Objective To observe the variations of intracellular localization and expression of importin 8 (IPO8) during
osteoblast differentiation. Methods Alizarin red staining, immunocytochemistry and real-time PCR were employed to examine
the changes in the intracellular localization and expression of IPO8 mRNA during induced osteogenic differentiation of human
osteoblast-like SaOS-2 cells. Results Numerous red mineralized nodules were observed on day 10 in the induced cells with
alizarin red staining. Immunocytochemical staining showed that IPO8 immunoreactivity was the strongest in the perinuclear
cytoplasm of the cells. On day 3 of osteoblast differentiation, IPO8 immunoreacivity in the cell nuclei became stronger. On day
7, IPO8 was located mainly in the nuclei, and on day 10 the cells were osteocyte-like and IPO8 was distributed in the
cytoplasm. Real-time PCR showed a significantly increased expression of OPN mRNA during osteoblast differentiation, and
the expression level of IPO8 mRNA was the highest on day 3 and declined on days 7 and 10. Conclusion The intracellular
localization and expression level of IPO8 undergo significant changes during osteogenesis, indicating its role in regulating
osteoblast differentiation.