全反式维甲酸诱导小鼠间充质干细胞C3H10T1/2表达碱性磷酸酶(1)
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【摘要】 目的 分析全反式维甲酸对小鼠间充质干细胞C3H10T1/2表达碱性磷酸酶的影响。方法 全反式维甲酸(10-6M)、维甲酸核受体RAR的激动剂Ch55与MAPK信号通路的阻滞剂处理C3H10T1/2细胞,钙钴染色检测碱性磷酸酶活性,定量RT-PCR方法检测量效关系以及时相分布;采用萤光素酶报告基因检测碱性磷酸酶的启动子是否接受维甲酸的调控。结果 维甲酸处理C3H10T1/2细胞24 h即能够明显上调ALP的活性,具有明显的剂量依赖特性,与RAR有关,无MAPK信号通路的参与,在其转录起始位点上游-512bp的范围内检测到核受体RAR的调控位点。结论 维甲酸能够通过核受体RAR直接上调碱性磷酸酶的转录水平。
【关键词】 全反式维甲酸;间充质干细胞;碱性磷酸酶;启动子
All Trans Retinoic Acid Upregulates Alkaline Phosphatase in Mouse Mesenchymal Stem Cell C3H10T1/2
【Abstract】 Objective To investigate the effects of all trans retinoic acid (ATRA) on the transcription level of mouse embryonic mesenchymal stem cell C3H10T1/2.Methods C3H10T1/2 were treated with all trans retinoic acid(10-6M), retinoic acid receptor ( RAR) agonist Ch55 and MAPK signal pathway inhibitor, alkaline phosphatase(ALP)activity were monitored by ALP staining, ALP transcription level were analyzed by quantitative RT-PCR; Systemic promoter analysis based on Luciferase reporter assay was undertaken to search for ALP promoter region to which RAR binds ......
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