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Published on: September 25, 2011
DNA microarray analysis of vitamin D-induced gene expression in a human colon carcinoma cell line
Richard J Wood1, Laurie Tchack, Giana Angelo
1Mineral Bioavailability Laboratory, Jean Mayer United States Department of Agriculture Human Nutrition Research Center on Aging at Tufts University, Boston 02111, USA. richard.wood@tufts.edu
Abstract:
The full extent to which 1,25-dihydroxyvitamin D(3) affects gene expression in human intestinal epithelial cells is unknown. We used oligonucleotide arrays to catalog vitamin D-induced changes in gene expression in Caco-2 cells, a human colon carcinoma cell line. Five paired sets of Caco-2 cell cultures were subjected to either control conditions or 1,25-dihydroxyvitamin D (10(-7) mol/l x 24 h), and RNA was analyzed on an Affymetrix cDNA array containing 12,625 human sequences. Only 13 sequences representing 12 distinct genes exhibited statistically significant changes in expression of twofold or greater and were also called as "present" or "marginal" by the array-reading software in all five experiments. Genes regulated by 1,25-dihydroxyvitamin D included two previously known genes (25-hydroxyvitamin D-24-hydroxylase and amphiregulin) and 10 genes (sorcin, Gem, adaptin-gamma, TIG1, CEACAM6, carbonic anhydrase XII, junB, ceruloplasmin, and two unidentified sequences) that were novel. We tested and independently confirmed the effect of 1,25-dihydroxyvitamin D on 11 of these genes by RT-PCR. Increased protein expression was tested and confirmed in two of the novel 1,25-dihydroxyvitamin D-regulated genes, ceruloplasmin and sorcin. The known function of these genes suggests that many of them could be involved in the antiproliferative effects of 1,25-dihydroxyvitamin D3.
Insights
The active form of vitamin D, 1,25-dihydroxyvitamin D(3), influences gene expression in intestinal cells. This study identified 12 genes, including novel ones, regulated by vitamin D in Caco-2 cells.
Area of Science:
- Molecular Biology
- Genomics
- Cell Biology
Background:
- The precise impact of 1,25-dihydroxyvitamin D(3) on gene expression in human intestinal epithelial cells remains incompletely understood.
- Vitamin D plays crucial roles in cellular functions, including proliferation and differentiation, particularly within the intestinal epithelium.
Purpose of the Study:
- To comprehensively catalog vitamin D-induced alterations in gene expression within a human colon carcinoma cell line (Caco-2).
- To identify novel genes regulated by 1,25-dihydroxyvitamin D(3) in intestinal cells and explore their potential roles.
Main Methods:
- Utilized oligonucleotide arrays (Affymetrix cDNA array with 12,625 human sequences) to analyze gene expression changes.
- Compared gene expression profiles in Caco-2 cells treated with 1,25-dihydroxyvitamin D(3) versus control conditions.
- Employed RT-PCR to validate changes in expression for selected genes and confirmed protein expression for novel targets.
Main Results:
- Identified 13 sequences (12 distinct genes) with statistically significant expression changes (≥ twofold) induced by 1,25-dihydroxyvitamin D(3).
- Confirmed regulation of previously known genes (25-hydroxyvitamin D-24-hydroxylase, amphiregulin) and identified 10 novel genes, including sorcin and ceruloplasmin.
- Validated vitamin D's effect on 11 genes via RT-PCR and confirmed increased protein levels for ceruloplasmin and sorcin.
Conclusions:
- 1,25-dihydroxyvitamin D(3) significantly impacts gene expression in human intestinal epithelial cells, regulating both known and novel genes.
- The identified genes, particularly ceruloplasmin and sorcin, may mediate the antiproliferative effects of 1,25-dihydroxyvitamin D(3) in the colon.
- This study provides a foundational catalog of vitamin D-responsive genes in the intestinal context, opening avenues for further research into vitamin D's biological functions.
