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1,25-dihydroxyvitamin D3 up-regulates expression of hsa-let-7a-2 through the interaction of VDR/VDRE in human lung cancer A549 cells.


Gene. 2013 Apr 5;


Authors: Guan H, Liu C, Chen Z, Wang L, Li C, Zhao J, Yu Y, Zhang P, Chen W, Jiang A


Abstract

AIMS: We aim to investigate the relationship between 1,25-(OH)2VD3 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)2VD3 on the expression of hsa-let-7a-2 after A549 cells were treated with 1,25-(OH)2VD3 (10(-8)~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) locates 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)2VD3 activating vitamin D receptor (VDR) could bind to this VDRE to promote hsa-let-7a-2 expression. RESULTS: We found that 1,25-(OH)2VD3 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)2VD3/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)2VD3. CONCLUSIONS: our data indicated that 1,25-(OH)2VD3 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)2VD3 might mediate the anti-proliferation effects of 1,25-(OH)2VD3 in lung cancer cells.

PMID: 23566834 [PubMed - as supplied by publisher]