Establishment and characterization of a stable cell line to evaluate cellular Runx2 activity
Establishment and characterization of a stable cell line to evaluate cellular Runx2 activity
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DOI：
10.1002/jcb.20038
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发表时间：
2004-04-15
影响因子：
4
通讯作者：
Ryoo, HM
中科院分区：
文献类型：
--
作者：
Kim, HJ;Park, HD;Ryoo, HM
Runx2 是成骨细胞从早期定型步骤到最终分化的重要转录因子。基于其在成骨细胞分化中的关键作用，Runx2 蛋白的转录活性比任何其他参数（例如 Runx2 mRNA 或蛋白水平）对于成骨细胞分化意味着更有价值的信息。因此，长期以来人们一直期望有一种灵敏、特异且一致的方法来确定 Runx2 转录活性。在这里，我们建议携带 6xOSE2-荧光素酶报告载体的稳定细胞系将是确定 Runx2 生物转录活性的良好评估系统。克隆细胞系的增殖率、细胞形状和成肌分化潜能与亲本前成肌细胞C2C12细胞相似。这些细胞对 Runx2 调节剂（例如 FGF2）有特异性反应。稳定细胞系的反应比用 Runx2 瞬时转染的细胞灵敏 5-6 倍。尽管任何 Runx 基因的过度表达都会刺激荧光素酶活性，但 Runx2 增强报告基因活性最高。总的来说，6xOSE2-luc 稳定细胞将是一个很好的生物学评估系统，用于评估细胞外 Runx2 调节刺激的活性以及刺激中涉及的信号转导途径。(C) 2004 Wiley-Liss, Inc.
Runx2 is an essential transcription factor for osteoblast differentiation from early commitment step to final differentiation. Based on its crucial role in osteoblast differentiation, the transcriptional activity of Runx2 protein implies more valuable information for osteoblast differentiation than any other parameters, such as Runx2 mRNA or protein level. Thus, a sensitive, specific, and consistent method to determine the Runx2 transcriptional activity has long been expected. Here we suggest a stable cell line that carries 6xOSE2-Luciferase reporter vector would be a good evaluation system to determine biological Runx2 transcriptional activity. The proliferation rate, cell shape, and the myogenic differentiation potential of the cloned cell line were similar to those of parental premyoblastic C2C12 cells. The cells specifically responded to Runx2 modulating agent such as FGF2. The stable cell line responded 5-6 folds more sensitively than the transiently transfected cells with Runx2. Though overexpression of any Runx gene stimulated the luciferase activity, Runx2 enhanced the reporter activity the highest. Collectively, the 6xOSE2-luc stable cells would be a good biological evaluation system to assess the activity of extracellular Runx2 modulating stimulations as well as the signal transduction pathways involved in the stimulations.(C) 2004 Wiley-Liss, Inc.