1,25-Dihydroxyvitamin D3 up-regulates expression of hsa-let-7a-2 through the interaction of VDR/VDRE in human lung cancer A549 cells
1,25-Dihydroxyvitamin D3 up-regulates expression of hsa-let-7a-2 through the interaction of VDR/VDRE in human lung cancer A549 cells
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DOI：
10.1016/j.gene.2013.03.065
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发表时间：
2013-06-15
期刊：
影响因子：
3.5
通讯作者：
Jiang, Anli
中科院分区：
文献类型：
--
作者：
Guan, Hengyun;Liu, Chunyan;Jiang, Anli
目的：本研究旨在探讨1，25-（OH）（2）VD 3在肺癌A549细胞中与hsa-let-7a的关系。采用荧光素酶报告基因和实时荧光定量PCR检测1，25-（OH）（2）VD 3对A549细胞hsa-let-7a-2表达的影响。25-（OH）（2）VD 3（10（-8）与10（-6）mol/L相似）。对pre-let-7a-2的5.0 Kb上游序列的分析表明，一个维生素D反应元件（VDRE）位于pre-let-7a-2的-2066/-2042 bp处。结果：1，25-（OH）（2）VD 3能以剂量依赖方式上调hsa-let-7a-2的表达。EMSA和ChIP结果表明，1，25-（OH）（2）VD 3/VDR可与pre-let-7a-2上游的VDRE相互作用。荧光素酶报告基因分析表明，该VDRE是一个功能性顺式元件，介导1，25-（OH）（2）VD 3上调hsa-let-7a-2的表达。我们的数据表明，1，25-（OH）（2）VD 3可上调肺癌细胞hsa-let-7a-2的转录，25-（OH）（2）VD 3可能介导了1，25-（OH）（2）VD 3对肺癌细胞的抗增殖作用。(C)2013爱思唯尔有限公司版权所有。
Aims: We aim to investigate the relationship between 1,25-(OH)(2)VD3 and hsa-let-7a in lung cancer A549 cells.Methods: Real-time PCR and luciferase reporter assays were used to detect the influence of 1,25-(OH)(2)VD3 on the expression of hsa-let-7a-2 after A549 cells were treated with 1,25-(OH)(2)VD3 (10(-8) similar to 10(-6) mol/L). Analysis of the 5.0 Kb upstream sequence of the pre-let-7a-2 showed that one vitamin D response element (VDRE) is located in -2066/-2042 bp of pre-let-7a-2. Electrophoretic mobility shift assays (EMSA), chromatin immunoprecipitation (ChIP) and luciferase reporter assays were performed to determine whether 1,25-(OH)(2)VD3 activating vitamin D receptor (VDR) could bind to this VDRE to promote hsa-let-7a-2 expression.Results: We found that 1,25-(OH)(2)VD3 could up-regulate the expression of hsa-let-7a-2 in a dose-dependent manner. The results of EMSA and ChIP demonstrated that 1,25-(OH)(2)VD3/VDR could interact with the VDRE in the upstream of pre-let-7a-2. Luciferase reporter assay showed that this VDRE is a functional cis-element mediating the up-regulation of hsa-let-7a-2 expression induced by 1,25-(OH)(2)VD3.Conclusions: Our data indicated that 1,25-(OH)(2)VD3 could up-regulate the transcription of hsa-let-7a-2 in lung cancer cells, and the up-regulation of hsa-let-7a-2 expression induced by 1,25-(OH)(2)VD3 might mediate the anti-proliferation effects of 1,25-(OH)(2)VD3 in lung cancer cells. (C) 2013 Elsevier B.V. All rights reserved.