Vitamin D co-regulates cell proliferation, differentiation and apoptosis in numerous tissues, including cancers. The known anti-proliferative and pro-apoptotic actions of the active metabolite of vitamin D,1,25-dihydroxy-vitamin D [1,25（OH）2 D] are mediated through binding to the vitamin D receptor（VDR）. Here,we report on the unexpected finding that stable knockdown of VDR expression in the human breast and prostate cancer cell lines, MDA-MB-231 and PC3, strongly induces cell apoptosis and inhib... 

【文章來源】：Bone Research. 2017,5(03)SCICSCD

【文章頁數(shù)】：12 頁

【文章目錄】：
INTRODUCTION
MATERIALS AND METHODS
    Cell culture
    Knockdown of VDR expression in cancer cells
    Cell growth assays
    Cell apoptosis assay
    Transcriptome analysis and Quantitative RT-PCR
    Western blot analysis
    Animal models
    Histological staining and immunohistochemistry
    Micro-computed tomography imaging
    Bone histomorphometry
    Statistical analysis
RESULTS
    Knockdown of VDR expression in MDA-MB-231 cells
    VDR knockdown in MDA-MB-231 cells reduces cellproliferation and increases cell apoptosis:in vitro studies As 1, 25 (OH) 2D3treatment suppresses cancer cell growth in vitro, and vitamin D deficiency enhances breast and prostate cancer growth in animal models of bonemetastasis, 8, 10, 13we hypothesized that ablation of the VDR in human breast cancer cells would promote tumor growth.As expected, 25treatment of VDR-expressing MDA-NT cells with 10-8mol·L-11, 25 (OH) 2D3significantly reduced in vitro cell growth (Figure 1d-f) and induced a 2-fold increase in apoptosis compared to MDA-NT cells (Figure 1g) .Surprisingly, however, when cultured under ligand-free conditions the growth of MDA-VDR-KD cel s was also significantly reduced, with growth curves similar to those of 1, 25 (OH) 2D3treated MDA-NT cells.As expected, treatment of VDR knockdown cells with 1, 25 (OH) 2D3had no further effect on growth (Figure 1d) .Compared to NT controls, cell proliferation was significantly reduced in MDA-VDR-KD cells, again of similar magnitude to that seen in1, 25 (OH) 2D3-treated MDA-NT cells (Figure 1e and f) .In contrast, VDR knockdown was associated with a pronounced rise in cel apoptosis:Thus, the proportion of TUNEL-positive cells was 12-fold higher in MDA-VDR-KD cells compared to MDA-NT controls (Figure 1g) .In keeping with these observations, Caspase 3 m RNA expression and cleaved Caspase 3 protein levels were significantly higher in MDA-VDR-KD than NT cells (Figure 1h and i) .
    VDR knockdown in MDA-MB-231 cells reduces cell growth:in vivo studies
    Knockdown of VDR in MDA-MB-231 cells reduces the Wnt/β-catenin signaling
DISCUSSION



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