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• W2006565377 abstract "Recent studies suggest that growth inhibition by 1,25-dihydroxyvitamin D3 represents an innovative approach to ovarian cancer therapy. To understand the molecular mechanism of 1,25-dihydroxyvitamin D3 action, we profiled the hormone-induced changes in the transcriptome of ovarian cancer cells using microarray technology. More than 200 genes were identified to be regulated by 1,25-dihydroxyvitamin D3. Reverse transcription-PCR analyses confirmed the regulation of a group of apoptosis-related genes, including the up-regulation of the decoy receptor that inhibits tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) action, TRAIL receptor 4, and the down-regulation of Fas, the receptor that mediates the action of Fas ligand. The regulation was further confirmed at the protein level. Consistent with the regulation of the death receptors, pretreatment with 1,25-dihydroxyvitamin D3 decreased apoptosis induced by TRAIL and Fas ligand. Because persistent 1,25-dihydroxyvitamin D3 treatment has been shown to induce apoptosis in ovarian cancer, the hormone appears to exert a dual effect on the death of ovarian cancer cells. Knockdown of TRAIL receptor 4 by RNA interference or ectopic expression of Fas relieved the suppressive effect of 1,25-dihydroxyvitamin D3, showing that molecular manipulation of death receptors is a viable approach to overcome the protective effect of 1,25-dihydroxyvitamin D3 on the apoptosis of ovarian cancer. These strategies may allow ovarian cancer patients to benefit from therapy with both 1,25-dihydroxyvitamin D3 and ligands for death receptors, such as TRAIL, shown to selectively induce apoptosis in cancer but not normal cells. Recent studies suggest that growth inhibition by 1,25-dihydroxyvitamin D3 represents an innovative approach to ovarian cancer therapy. To understand the molecular mechanism of 1,25-dihydroxyvitamin D3 action, we profiled the hormone-induced changes in the transcriptome of ovarian cancer cells using microarray technology. More than 200 genes were identified to be regulated by 1,25-dihydroxyvitamin D3. Reverse transcription-PCR analyses confirmed the regulation of a group of apoptosis-related genes, including the up-regulation of the decoy receptor that inhibits tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) action, TRAIL receptor 4, and the down-regulation of Fas, the receptor that mediates the action of Fas ligand. The regulation was further confirmed at the protein level. Consistent with the regulation of the death receptors, pretreatment with 1,25-dihydroxyvitamin D3 decreased apoptosis induced by TRAIL and Fas ligand. Because persistent 1,25-dihydroxyvitamin D3 treatment has been shown to induce apoptosis in ovarian cancer, the hormone appears to exert a dual effect on the death of ovarian cancer cells. Knockdown of TRAIL receptor 4 by RNA interference or ectopic expression of Fas relieved the suppressive effect of 1,25-dihydroxyvitamin D3, showing that molecular manipulation of death receptors is a viable approach to overcome the protective effect of 1,25-dihydroxyvitamin D3 on the apoptosis of ovarian cancer. These strategies may allow ovarian cancer patients to benefit from therapy with both 1,25-dihydroxyvitamin D3 and ligands for death receptors, such as TRAIL, shown to selectively induce apoptosis in cancer but not normal cells. Vitamin D3 (VD) 2The abbreviations used are: VDvitamin D31,25-(OH)2D31,25-dihydroxyvitamin D3OCaovarian cancerTRAILtumor necrosis factor-related apoptosis-inducing ligandTRAIL-R2TRAIL receptor 2TRAIL-R4TRAIL receptor 4FasLFas ligandSAMsignificance analysis of microarrayRTreverse transcriptionMTTmethylthiazole tetrazoliumGFPgreen fluorescence proteinEGFPenhanced green fluorescence proteinsiRNAsmall interference RNAAMC7-amino-4-methylcoumarin. is a steroid hormone best known for its role in calcium homeostasis. Its deficiency causes rickets in children and osteomalacia in adults. VD also regulates the proliferation and differentiation of normal and malignant cells of many tissue types. Because sunlight controls the first step of VD synthesis, namely, the photoconversion of 7-dehydrocholesterol to vitamin D3 (1Webb A.R. Kline L. Holick M.F. J. Clin. Endocrinol. Metab. 1988; 67: 373-378Crossref PubMed Scopus (1374) Google Scholar), the hormone is considered a “sun” medicine that is effective for the treatment and/or prevention of type II rickets, osteoporosis, autoimmune disorders, as well as epithelial cancers of many types. vitamin D3 1,25-dihydroxyvitamin D3 ovarian cancer tumor necrosis factor-related apoptosis-inducing ligand TRAIL receptor 2 TRAIL receptor 4 Fas ligand significance analysis of microarray reverse transcription methylthiazole tetrazolium green fluorescence protein enhanced green fluorescence protein small interference RNA 7-amino-4-methylcoumarin. Ovarian cancer (OCa) is a fatal disease with an overall 5-year survival rate of about 40%. OCa mortality and incidence rates are lower in countries within 20 degrees of the equator (2Muir C. Waterhouse J. Mack T. Powell J. Whelan S. Cancer Incidence in Five Continents, International Agency for Research on Cancer Control (IARC). 1987; V (Scientific Publication No. 88, Vol. , IARC, Lyon, France)Google Scholar) where there is a high amount of sunlight. In the United States, women between the ages of 45 and 54 living in the North have 5 times the OCa mortality rate of women living in Southern states (3Devesa S.S. Grauman M.A. Blot W.J. Pennello G.A. Hoover R.N. Fraumeni Jr., J.F. Atlas of Cancer Mortality in the United States, 1950–1994. 1999; (National Institutes of Health Publication No. 99–4564, NCI, National Institutes of Health, Bethesda, MD)Google Scholar). The inverse correlation between sunlight exposure and OCa mortality indicates that decreased synthesis of VD may contribute to OCa initiation and/or progression. This idea has been further substantiated by recent studies (4Jiang F. Li P. Fornace Jr., A.J. Nicosia S.V. Bai W. J. Biol. Chem. 2003; 278: 48030-48040Abstract Full Text Full Text PDF PubMed Scopus (120) Google Scholar, 5Miettinen S. Ahonen M.H. Lou Y.R. Manninen T. Tuohimaa P. Syvala H. Ylikomi T. Int. J. Cancer. 2004; 108: 367-373Crossref PubMed Scopus (40) Google Scholar, 6Li P. Li C. Zhao X. Zhang X. Nicosia S.V. Bai W. J. Biol. Chem. 2004; 279: 25260-25267Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar, 7Zhang X. Jiang F. Li P. Li C. Ma Q. Nicosia S.V. Bai W. Clin. Cancer Res. 2005; 11: 323-328PubMed Google Scholar) showing that 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3), the active form of VD, inhibits the growth of multiple OCa cell lines (4Jiang F. Li P. Fornace Jr., A.J. Nicosia S.V. Bai W. J. Biol. Chem. 2003; 278: 48030-48040Abstract Full Text Full Text PDF PubMed Scopus (120) Google Scholar, 5Miettinen S. Ahonen M.H. Lou Y.R. Manninen T. Tuohimaa P. Syvala H. Ylikomi T. Int. J. Cancer. 2004; 108: 367-373Crossref PubMed Scopus (40) Google Scholar, 6Li P. Li C. Zhao X. Zhang X. Nicosia S.V. Bai W. J. Biol. Chem. 2004; 279: 25260-25267Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar) and OVCAR3 tumor xenografts in nude mice (7Zhang X. Jiang F. Li P. Li C. Ma Q. Nicosia S.V. Bai W. Clin. Cancer Res. 2005; 11: 323-328PubMed Google Scholar). Effects of VD are mediated through the vitamin D receptor (8Baker A.R. McDonnell D.P. Hughes M. Crisp T.M. Mangelsdorf D.J. Haussler M.R. Pike J.W. Shine J. O'Malley B.W. Proc. Natl. Acad. Sci. U. S. A. 1988; 85: 3294-3298Crossref PubMed Scopus (878) Google Scholar), which is a member of the steroid/thyroid receptor superfamily of ligand-regulated transcription factors. In response to the activation by 1,25-(OH)2D3, the receptor recruits multiple co-activators, including members of the p160 SRC family (9Onate S.A. Tsai S.Y. Tsai M.J. O'Malley B.W. Science. 1995; 270: 1354-1357Crossref PubMed Scopus (2063) Google Scholar), which are associated with histone acetyltransferase activity, and the DRIP complex (10Rachez C. Freedman L.P. Gene (Amst.). 2000; 246: 9-21Crossref PubMed Scopus (290) Google Scholar), which serves as a mediator between the vitamin D receptor and RNA polymerase II complex. Recent studies (11Hedlund T.E. Moffatt K.A. Miller G.J. Endocrinology. 1996; 137: 1554-1561Crossref PubMed Google Scholar) have suggested that the anti-tumor activity of VD is mediated through its effect on gene transcription. In OCa cells, we have shown that the transcriptional up-regulation of GADD45 is essential for the 1,25-(OH)2D3-induced cell cycle arrest at the G2/M checkpoint (4Jiang F. Li P. Fornace Jr., A.J. Nicosia S.V. Bai W. J. Biol. Chem. 2003; 278: 48030-48040Abstract Full Text Full Text PDF PubMed Scopus (120) Google Scholar). 1,25-(OH)2D3 also increases p27 protein stability in OCa cells through down-regulation of Skp2 and cyclin E mRNA to induce cell cycle arrest at the G1/S checkpoint (6Li P. Li C. Zhao X. Zhang X. Nicosia S.V. Bai W. J. Biol. Chem. 2004; 279: 25260-25267Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar). These studies suggest that the action of 1,25-(OH)2D3 through the nuclear activation of vitamin D receptor is important for the anti-tumor activity of the hormone in OCa. Because the anti-tumor activity of 1,25-(OH)2D3 is mediated through regulation of gene expression, a more complete understanding of the mechanism underlying 1,25-(OH)2D3 action in OCa cells necessitates identification of changes in gene expression induced by the hormone. In the present study, we profile transcriptional changes in OVCAR3 cells induced by 1,25-(OH)2D3 using multiple independent microarray analyses. We concentrated our subsequent analysis to a few apoptosis-related genes from this list. Our studies reveal that 1,25-(OH)2D3 regulates the expression of death receptors and protects cancer cells from apoptosis induced by death ligands. The findings suggest that a better understanding of both beneficial and adverse effects of 1,25-(OH)2D3 in OCa cells is necessary for the development of effective OCa therapeutic strategies that utilize active VD compounds, either alone or in combination with other apoptosis-inducing agents. Chemical Reagents, Antibodies, and Cell Cultures—1,25-(OH)2D3 was purchased from Calbiochem (La Jolla, CA). TRAIL, biological active Fas ligand, staurosporine, calcium ionophore A23187, and etoposide were from Sigma. The antibodies anti-TRAIL (BD Pharmingen), anti-TRAIL receptor 4 (TRAIL-R4), anti-Fas (Santa Cruz Biotechnology Inc., Santa Cruz, CA), anti-TRAIL-R2 (Oncogene Research Products, San Diego, CA), and anti-caspase-7 (Cell Signaling Technology Inc., Beverly, MA) were purchased commercially. All cell lines used in the study have been described previously (6Li P. Li C. Zhao X. Zhang X. Nicosia S.V. Bai W. J. Biol. Chem. 2004; 279: 25260-25267Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar). OVCAR3 cells were cultured in RPMI 1640 medium supplemented with 15% fetal bovine serum, 2 mm l-glutamine, penicillin (50 units/ml), streptomycin (50 μg/ml), 10 mm HEPES, 1 mm sodium pyruvate, 4.5 g/liter glucose, 1.5 g/liter sodium bicarbonate, and 10 μg/ml bovine insulin. Other cell lines were maintained in Dulbecco's modified Eagle's medium (Invitrogen) containing 10% fetal bovine serum. For 1,25-(OH)2D3 treatment, the hormone was dissolved in ethanol, diluted to the desired concentration in culturing medium, and added to the cells. For longer treatments, the medium was replaced with fresh medium containing 1,25-(OH)2D3 or vehicle every third day. Microarray Analyses and Data Statistics—For microarray analyses, OVCAR3 cells at about 70% confluence were treated with 10-7 m 1,25-(OH)2D3 for 0, 8, 24, or 72 h. 1,25-(OH)2D3 was added at different times to allow the treated cells to be harvested at the same time. Ethanol was included as vehicle controls, and all cells were exposed to the same amount of ethanol for the same length of time. Total RNA was extracted using TRIzol reagent (Invitrogen) and further purified using Qiagen RNeasy columns. An initial experiment was performed using the U95Av2 chip from Affymetrix Inc. (Santa Clara, CA). From this experiment it was determined that GADD45 expression changed in response to VD (4Jiang F. Li P. Fornace Jr., A.J. Nicosia S.V. Bai W. J. Biol. Chem. 2003; 278: 48030-48040Abstract Full Text Full Text PDF PubMed Scopus (120) Google Scholar). Three subsequent experiments were performed using the U133A chips (Affymetrix Inc.). In these experiments the response of the cells to 1,25-(OH)2D3 was determined by Northern blotting analysis of GADD45 before using the isolated RNA for microarray analysis. The RNA samples were processed for hybridization to microarrays by the Microarray Core Facility of the H. Lee Moffitt Cancer Center. Briefly, RNA was converted to double-stranded cDNA using an oligo(dT)24 primer containing a T7 promoter sequence. The resulting double-stranded cDNA was transcribed into biotin-labeled cRNA using T7 RNA polymerase. This cRNA was hybridized to GeneChip probe arrays. Unbound RNA was washed from the chips and the remaining biotinylated RNA was stained and the chips scanned. Scanned chip images were analyzed using microarray suite software (MAS 5.0) from Affymetrix. In total, four independent experiments were carried out. Raw data were processed using Irizarry and Speed's Robust Multichip Analysis. After normalization, significance analysis of microarray (SAM) analysis (12Tusher V.G. Tibshirani R. Chu G. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 5116-5121Crossref PubMed Scopus (9802) Google Scholar) was performed by comparing gene expression profiles at time points of 8, 24, and 72 h to those of vehicle control (0 time point). Significant genes were clustered using GeneSpring software 6.0 (Silicon Genetics, Redwood City, CA). This resulted in groupings based on the general pattern of expression changes with time following 1,25-(OH)2D3 addition. The final list of genes was then generated by evaluating each probeset from the SAM analysis for consistency. A gene was considered to be consistently regulated if the same gene expression profile was obtained in all three experiments performed with the U133A chips and detected by all probesets. Gene identification was based on the sequence of the probes used on the arrays (13Harbig J. Sprinkle R. Enkemann S.A. Nucleic Acids Res. 2005; 33: e31Crossref PubMed Scopus (102) Google Scholar). Reverse Transcription Polymerase Chain Reaction (RT-PCR)— RT-PCR was conducted according to the protocol of Invitrogen with minor modifications. Briefly, 1 μg of pooled RNA from three independent experiments was reverse transcribed with the oligo(dT)20 primer in a total reaction volume of 20 μl. The amplification of target genes was carried out using 1 μl of reverse transcription product in a total reaction volume of 25 μl. PCR was performed using a GeneAmp PCR System 2700 (Applied Biosystem, Inc., Foster City, CA) for about 25 cycles with each cycle consisting of denaturing at 94 °C for 30 s followed by annealing for 45 s at various temperatures (supplemental Table S1) and elongation for 1 min at 72 °C. PCR products were visualized by electrophoresis on 1.5% agarose gels. Primers were synthesized by Invitrogen and the sequences are listed in supplemental Table S1. Immunoblotting Analysis, Cell Growth, and Apoptotic Assays—For immunoblotting analyses, cellular extracts were separated on SDS-PAGE gels and blotted onto nitrocellulose membranes. Antibodies were used at 1:500 to 1:1000 dilutions. Proteins were detected using ECL as described (6Li P. Li C. Zhao X. Zhang X. Nicosia S.V. Bai W. J. Biol. Chem. 2004; 279: 25260-25267Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar). To assay cell growth, methylthiazole tetrazolium (MTT) assays were performed as described (6Li P. Li C. Zhao X. Zhang X. Nicosia S.V. Bai W. J. Biol. Chem. 2004; 279: 25260-25267Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar). For each data point, eight samples were analyzed in parallel. Absorption at 595 nm (A595 nm) was measured on a MRX microplate reader (DYNEX Technologies, Chantilly, VA). The apoptotic index of cells treated with death ligands and 1,25-(OH)2D3 was determined by flow cytometry following staining with Annexin V-fluorescein isothiocyanate and propidium iodide according to the manufacturer's protocol (Annexin V kit, Santa Cruz Biotechnology Inc.). Flow cytometry was performed in a BD Biosciences FACS-Calibur flow cytometer. The data from 10,000 cells per time point were acquired and analyzed using CellQuest software. The apoptotic index of transfected cells was determined as previously described (6Li P. Li C. Zhao X. Zhang X. Nicosia S.V. Bai W. J. Biol. Chem. 2004; 279: 25260-25267Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar). Transfected cells were fixed in 3.7% formaldehyde/phosphate-buffered saline and stained with 4′,6′-diamidino-2-phenylindole. The apoptotic index of green fluorescence protein (GFP)-positive cells was determined by scoring 400 GFP-positive cells in randomly selected microscopic fields for chromatin condensation and apoptotic body formation. Caspase Assays—The activity of caspase-3 was determined by the amount of fluorescent AMC liberated from the cleavage of the peptide substrate, Ac-DEVD-AMC, specific for caspase-3, using an assay kit from BD Biosciences. AMC was measured using a spectrofluorometer at excitation of 380 nm and emission of 420 nm. Caspase-7 activity was similarly determined by the amount of free fluorescent AFC released from substrate, Ac-DEVD-AFC, by caspase-7 immunoprecipitates following the protocol of the assay kit from Cell Signaling (Beverly, MA). AFC fluorescence was measured at excitation of 400 nm and emission of 505 nm. Plasmid, siRNA, and Transfection—TRAIL-R4 specific siRNA was synthesized from Upstate Inc. (Chicago, IL). The sense and antisense sequences were 5′-GGACAUGCAAAGGAAACAAtt-3′ and 5′-UUGUUUCCUUUGCAUGUCCtt-3′, respectively. OVCAR3 cells were transfected with either pCMV-Fas and pEGFP or TRAIL-R4 siRNA using Lipofectamine or Oligofectamine according to the manufacturer's protocol (Invitrogen). pCMV plasmid and nonspecific siRNA were used as controls. Cells were pretreated with 10-7 m 1,25-(OH)2D3 or vehicle for 3 days before the transfection, then 6 h post-transfection, cells were replaced in fresh medium containing 1,25-(OH)2D3 or vehicle. Forty-eight h later, cells were processed for immunoblotting analyses or treated with TRAIL or Fas ligand for apoptosis assays. Identification of 1,25-(OH)2D3-regulated Genes by Microarray in OVCAR3 Cells—To profile the changes in the pattern of gene expression in human OCa cells induced by 1,25-(OH)2D3, OVCAR3 cells were treated with 10-7 m 1,25-(OH)2D3 for 0, 8, 24, or 72 h in four independent experiments. Total RNA was isolated, reverse transcribed, and subjected to microarray analyses. The first analysis was performed with the U95Av2 Affymetrix chips, which identified GADD45 among many others as being up-regulated. Subsequent analysis with conventional approaches confirmed this and identified GADD45 as a primary target gene for 1,25-(OH)2D3 (4Jiang F. Li P. Fornace Jr., A.J. Nicosia S.V. Bai W. J. Biol. Chem. 2003; 278: 48030-48040Abstract Full Text Full Text PDF PubMed Scopus (120) Google Scholar). To ensure the reproducibility of the microarray data, three independent analyses were subsequently performed with the U133A Affymetrix chips, which contain 22,283 probe sets and can be used to analyze the expression level of ∼15,000 known human genes. For quality control, RNA samples were first tested by Northern blots for up-regulation of GADD45 to ensure that the cells from which the RNA was isolated had responded to the treatment (data not shown). Samples from cells treated with the hormone for 3 days were included to ensure that genes that are regulated by persistent 1,25-(OH)2D3 treatment might be identified. These genes could mediate the anti-tumor activity of the hormone even though they may not be primary target genes. The raw data from the three microarray studies with U133A were first analyzed with SAM (12Tusher V.G. Tibshirani R. Chu G. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 5116-5121Crossref PubMed Scopus (9802) Google Scholar), a statistical method based on the conventional t test but developed specifically for microarray analysis. Using the 5% false discovery rate, we found that 710 probesets representing about 500 genes were modulated by 1,25-(OH)2D3. We reduced this list by requiring that a gene must change by at least 2.5-fold at one point during the time course of the experiment. The selected genes were clustered into groups using GeneSpring software, which generated five clusters (Fig. 1), of which, clusters 1 to 3 were up-regulated and clusters 4 and 5 were down-regulated. Genes in clusters 1 and 4 were induced or repressed at 8- or 24-h treatment and prolonged treatment did not change the fold regulation dramatically. Examples are GADD45 and 1,25-(OH)2D3 24-hydroxylase genes, which both contain vitamin D response elements in their genome (4Jiang F. Li P. Fornace Jr., A.J. Nicosia S.V. Bai W. J. Biol. Chem. 2003; 278: 48030-48040Abstract Full Text Full Text PDF PubMed Scopus (120) Google Scholar, 14Zou A. Elgort M.G. Allegretto E.A. J. Biol. Chem. 1997; 272: 19027-19034Abstract Full Text Full Text PDF PubMed Scopus (78) Google Scholar). Genes in clusters 2 and 5 were steadily up- or down-regulated and their fold regulation kept increasing as the time of treatment was extended. The cyclin A1 gene fell into cluster 3, which was repressed significantly at 8 h treatment but became induced at 72 h (2.58-fold) of 1,25-(OH)2D3 treatment. The final gene list contains only those genes whose pattern of expression was consistent in all three experiments and across all probesets that detect the same gene. This reduced the list to 58 genes that appeared to be up-regulated (TABLE ONE) and 38 genes that appeared to be down-regulated (TABLE TWO). Many of these genes were also identified in the previous experiment using U95Av2 chips (TABLES ONE and TWO).TABLE ONEGenes up-regulated by 1,25-(OH)2D3 (total: 58) Genes are grouped by gene molecular functions. Mean -fold changes and S.D. are presented.GenBankGene name-Fold change (mean ± S.D.)qaq value refers to the possibility of a false positive for a specific gene by SAM analysisU958 h24 h72 h%Cell cycle and apoptosisNM_001924GADD45bGenes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell types2.77 ± 0.093.71 ± 0.134.02 ± 0.152.6√NM_003914Cyclin A1bGenes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell types–1.63 ± 0.11–1.17 ± 0.212.58 ± 0.181.1√NM_003810TRAIL2.76 ± 0.022.69 ± 0.102.31 ± 0.171.5√NM_003840TRAIL-R41.85 ± 0.132.83 ± 0.443.00 ± 0.404.6√Oncogene and tumor suppressorsNM_004675Ras homolog gene family member I (ARHI)1.78 ± 0.442.43 ± 0.116.51 ± 0.251.2√NM_022337RAB38, Ras oncogene family–1.01 ± 0.202.01 ± 0.282.70 ± 0.191.6N/AcN/A, not applicableNM_005252Fos6.79 ± 0.649.42 ± 0.2310.61 ± 0.084.1√NM_004442EphB2–1.05 ± 0.302.98 ± 0.509.38 ± 0.471.6√NM_006408Anterior gradient 2 homolog2.77 ± 0.124.11 ± 0.277.00 ± 0.210.6N/AEnzymesNM_00078224-HydroxylasebGenes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell types46.8 ± 0.0554.94 ± 0.2455.91 ± 0.190.6√NM_002774Kallikrein 62.60 ± 0.095.56 ± 0.147.21 ± 0.180.6√NM_005308G protein-coupled receptor kinase 5 (GPRK5)bGenes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell types4.17 ± 0.258.80 ± 0.2713.34 ± 0.101.1√NM_007207Dual specificity phosphatase 10 (DUSP10)bGenes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell types2.92 ± 0.114.23 ± 0.083.67 ± 0.080.6√NM_015170Sulfatase 11.29 ± 0.142.64 ± 0.153.65 ± 0.160.9√NM_000402Glucose-6-phosphate dehydrogenase (G6PD)bGenes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell types1.41 ± 0.123.23 ± 0.115.01 ± 0.200.6√NM_003784Serine proteinase inhibitor clade B, 71.99 ± 0.5510.11 ± 0.2021.30 ± 0.080.6N/ANM_016931NADPH oxidase 4 (NOX4)1.27 ± 0.352.63 ± 0.253.21 ± 0.144.6N/ANM_002168Mitochondrial isocitrate dehydrogenase 2 (NADP+)1.05 ± 0.121.92 ± 0.132.50 ± 0.191.1√NM_002849Protein-tyrosine phosphatase, receptor type, RbGenes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell types1.50 ± 0.021.95 ± 0.272.66 ± 0.320.6N/ANM_012232RNA polymerase I and transcript release factor1.20 ± 0.162.22 ± 0.242.98 ± 0.081.1N/ANM_001056Sulfotransferase family, cytosolic, 1C, member 11.32 ± 0.422.48 ± 0.282.75 ± 0.094.6N/ANM_000904NAD(P)H dehydrogenase, quinone 21.49 ± 0.162.24 ± 0.092.95 ± 0.174.6N/ANM_006741Protein phosphatase 1, regulatory (inhibitor) subunit 1A1.53 ± 0.243.03 ± 0.272.80 ± 0.114.6N/ANM_001977Glutamyl aminopeptidase (aminopeptidase A)1.88 ± 0.273.41 ± 0.032.73 ± 0.181.5N/CdN/C, -fold change less than 2.50NM_006105Rap guanine nucleotide exchange factor (GEF) 32.58 ± 0.173.52 ± 0.052.69 ± 0.252.7N/ANM_001993Coagulation factor III1.61 ± 0.181.81 ± 0.182.51 ± 0.214.6N/ASignal transducers, growth factors, and receptorsNM_004626Wingless-type MMTV integration site, 11 (WNT11)1.60 ± 0.1512.15 ± 0.2432.85 ± 0.180.6√NM_000728Calcitonin-related polypeptide β3.13 ± 0.174.60 ± 0.276.40 ± 0.172.6√NM_001401Endothelial differentiation, lysophosphatidic acid G-protein-coupled receptor 2 (Edg2)2.00 ± 0.313.99 ± 0.435.28 ± 0.150.6√M37435Human macrophage-specific colony-stimulating factor1.43 ± 0.092.41 ± 0.062.50 ± 0.150.6√Transcription factorsNM_021784Forkhead box A21.66 ± 0.233.52 ± 0.253.24 ± 0.171.5N/ANM_005257GATA-63.37 ± 0.093.87 ± 0.174.31 ± 0.134.6√Immunity proteins, structural proteins, cell adhesion molecules, and cytoskeletonNM_004345Cathelicidin antimicrobial peptide10.72 ± 0.3228.19 ± 0.1035.89 ± 0.100.6N/ANM_001845Collagen, type IV, α11.85 ± 0.133.16 ± 0.123.08 ± 0.091.5√NM_001846Collagen, type IV, α21.46 ± 0.092.25 ± 0.032.78 ± 0.111.2√NM_001003407Actin binding LIM protein 1 (ABLIM)1.38 ± 0.013.17 ± 0.253.23 ± 0.210.9N/AAL832563Neural cell adhesion molecule 11.70 ± 0.212.64 ± 0.213.23 ± 0.174.0N/ANM_000582NephropontinbGenes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell types2.12 ± 0.342.74 ± 0.026.11 ± 0.063.3N/ANM_005554Keratin 6A1.02 ± 0.371.96 ± 0.324.27 ± 0.140.6√NM_001407Cadherin, EGF LAG seven-pass G-type receptor 3–1.18 ± 0.641.11 ± 0.883.05 ± 0.143.3N/ANM_005261GTP-binding protein overexpressed in skeletal muscle (GEM)bGenes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell types1.36 ± 0.081.97 ± 0.203.16 ± 0.212.6√NM_000138Fibrillin 1 (FBN1)1.16 ± 0.453.33 ± 0.525.38 ± 0.274.6√OthersXM_378901Filaggrin gene1.31 ± 0.042.42 ± 0.235.08 ± 0.120.6√NM_002888Retinoic acid receptor responder 1 (RARRES1)bGenes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell types3.04 ± 0.235.71 ± 0.219.73 ± 0.130.6√AK092245Clone 141H5 contains parts of a novel chordin-like protein1.46 ± 0.052.95 ± 0.063.81 ± 0.000.6√NM_017724Leucine-rich repeat interacting protein 21.53 ± 0.092.65 ± 0.263.28 ± 0.131.5N/ANM_018431Docking protein 52.08 ± 0.073.19 ± 0.033.26 ± 0.140.6√AF191019Estradiol-induced (E2IG4)2.63 ± 0.173.68 ± 0.023.44 ± 0.160.6N/ANM_017734Palmdelphin4.52 ± 0.126.51 ± 0.067.55 ± 0.280.6N/ANM_030915Likely ortholog of mouse limb-bud and heart gene1.96 ± 0.132.77 ± 0.073.08 ± 0.130.6N/AD84109RBP-MS/type 31.55 ± 0.091.79 ± 0.252.63 ± 0.100.3√NM_020182Transmembrane prostate androgen-induced RNA (TMEPAI)1.75 ± 0.576.08 ± 0.603.98 ± 0.104.6N/AAL049370cDNA DKFZp586D09181.07 ± 0.281.21 ± 0.212.51 ± 0.120.6N/CNM_134264WD repeat and SOCS box-containing 12.56 ± 0.251.82 ± 0.102.10 ± 0.370.6N/ANM_000039Apolipoprotein A-I1.30 ± 0.862.75 ± 0.263.96 ± 0.180.9N/ANM_138444Potassium channel tetramerization domain containing 122.53 ± 0.163.50 ± 0.103.78 ± 0.140.6N/ANM_003982Solute carrier family 7 (cationic amino acid transporter, y+ system), member 71.21 ± 0.132.14 ± 0.223.57 ± 0.424.0N/ANM_002343Lactotransferrin1.17 ± 0.092.83 ± 0.873.37 ± 0.442.2N/Aa q value refers to the possibility of a false positive for a specific gene by SAM analysisb Genes previously reported to be 1,25-(OH)2D3-regulated in microarray analyses of other cell typesc N/A, not applicabled N/C, -fold change less than 2.50 Open table in a new tab TABLE TWOGenes down-regulated by 1,25-(OH)2D3 (total: 38) Genes are grouped by gene molecular functions. Mean -fold changes and S.D. are presented.GenBankGene name-Fold change (mean ± S.D.)qaq value refers to the possibility of a false positive for a specific gene by SAM analysisU958 h24 h72 h%Cell cycle and apoptosisNM_014059Response gene to complement 32 (RGC 32)–4.90 ± 0.13–8.14 ± 0.38–5.47 ± 0.080.6N/AbN/A, not applicableNM_000043Fas–1.54 ± 0.14–1.96 ± 0.27–2.50 ± 0.210.6√Oncogene and tumor suppressorsNM_005564Lipocalin 2 (LCN2)–1.04 ± 0.13–1.59 ± 0.23–3.05 ± 0.210.6N/ANM_002089GRO-β–1.68 ± 0.20–3.92 ± 0.10–3.35 ± 0.250.6√NM_002090GRO-γ–1.69 ± 0.19–3.32 ± 0.16–2.54 ± 0.362.2√NM_007177TU3A–1.64 ± 0.10–3.35 ± 0.17–5.97 ± 0.250.6N/AEnzymesNM_001001567Phosphodiesterase 9A–1.69 ± 0.21–4.14 ± 0.33–6.82 ± 0.940.6√NM_001116Adenylate cyclase 9–1.47 ± 0.12–3.01 ± 0.42–3.60 ± 0.070.6√NM_000295Serine proteinase inhibitor, clade A, member 1–1.46 ± 0.26–1.68 ± 0.23–3.62 ± 0.333.3N/ASignal transducers, growth factors, and receptorsNM_004591Small inducible cytokine subfamily A (Cys-Cys), 20 (SCYA20)–2.09 ± 0.39–23.64 ± 0.31–14.59 ± 0.770.6N/ANM_002993Small inducible cytokine subfamily B (Cys-X-Cys), 6 (SCYB6)–1.53 ± 0.24–2.88 ± 0.12–2.82 ± 0.151.2N/ANM_000627Latent transforming growth factor β-binding protein 1–1.41 ± 0.22–5.36 ± 0.20–6.27 ± 0.380.6N/ANM_001999Fibrillin" @default.
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• W2006565377 date "2005-10-01" @default.
• W2006565377 modified "2023-10-16" @default.
• W2006565377 title "Suppression of Death Receptor-mediated Apoptosis by 1,25-Dihydroxyvitamin D3 Revealed by Microarray Analysis" @default.
• W2006565377 cites W1898285755 @default.
• W2006565377 cites W1967694316 @default.
• W2006565377 cites W1972104782 @default.
• W2006565377 cites W1973195685 @default.
• W2006565377 cites W1974280437 @default.
• W2006565377 cites W1984371642 @default.
• W2006565377 cites W1984380932 @default.
• W2006565377 cites W1984998883 @default.
• W2006565377 cites W1985812086 @default.
• W2006565377 cites W1991048381 @default.
• W2006565377 cites W1992934249 @default.
• W2006565377 cites W1994621911 @default.
• W2006565377 cites W1998878159 @default.
• W2006565377 cites W2000816728 @default.
• W2006565377 cites W2003697339 @default.
• W2006565377 cites W2015063563 @default.
• W2006565377 cites W2019122456 @default.
• W2006565377 cites W2026194974 @default.
• W2006565377 cites W2052375963 @default.
• W2006565377 cites W2052647271 @default.
• W2006565377 cites W2054630945 @default.
• W2006565377 cites W2055897952 @default.
• W2006565377 cites W2057773030 @default.
• W2006565377 cites W2061858357 @default.
• W2006565377 cites W2065893174 @default.
• W2006565377 cites W2068113812 @default.
• W2006565377 cites W2072419218 @default.
• W2006565377 cites W2077575239 @default.
• W2006565377 cites W2079716869 @default.
• W2006565377 cites W2092410657 @default.
• W2006565377 cites W2100639130 @default.
• W2006565377 cites W2101270985 @default.
• W2006565377 cites W2117263637 @default.
• W2006565377 cites W2130278329 @default.
• W2006565377 cites W2136706713 @default.
• W2006565377 cites W2143162174 @default.
• W2006565377 cites W2146940464 @default.
• W2006565377 cites W2148615450 @default.
• W2006565377 cites W2157043934 @default.
• W2006565377 cites W2157330336 @default.
• W2006565377 cites W2157735691 @default.
• W2006565377 cites W2157751466 @default.
• W2006565377 cites W2162741492 @default.
• W2006565377 cites W2197704460 @default.
• W2006565377 cites W2323027157 @default.
• W2006565377 cites W4294107304 @default.
• W2006565377 cites W82539334 @default.
• W2006565377 doi "https://doi.org/10.1074/jbc.m506648200" @default.
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