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Vitamin D(3) promotes the differentiation of colon carcinoma cells by the induction of E-cadherin and the inhibition of beta-catenin signaling.

Pálmer HG, González-Sancho JM, Espada J, Berciano MT, Puig I, Baulida J, Quintanilla M, Cano A, de Herreros AG, Lafarga M, Muñoz A - J. Cell Biol. (2001)

Bottom Line: Accordingly, 1alpha,25(OH)2D(3) repressed beta-catenin-TCF-4 transcriptional activity.Also, 1alpha,25(OH)2D(3) inhibited expression of beta-catenin-TCF-4-responsive genes, c-myc, peroxisome proliferator-activated receptor delta, Tcf-1, and CD44, whereas it induced expression of ZO-1.Our results show that 1alpha,25(OH)2D(3) induces E-cadherin and modulates beta-catenin-TCF-4 target genes in a manner opposite to that of beta-catenin, promoting the differentiation of colon carcinoma cells.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Investigaciones Biomédicas "Alberto Sols, " Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, E-28029 Madrid, Spain.

ABSTRACT
The beta-catenin signaling pathway is deregulated in nearly all colon cancers. Nonhypercalcemic vitamin D3 (1alpha,25-dehydroxyvitamin D(3)) analogues are candidate drugs to treat this neoplasia. We show that these compounds promote the differentiation of human colon carcinoma SW480 cells expressing vitamin D receptors (VDRs) (SW480-ADH) but not that of a malignant subline (SW480-R) or metastasic derivative (SW620) cells lacking VDR. 1alpha,25(OH)2D(3) induced the expression of E-cadherin and other adhesion proteins (occludin, Zonula occludens [ZO]-1, ZO-2, vinculin) and promoted the translocation of beta-catenin, plakoglobin, and ZO-1 from the nucleus to the plasma membrane. Ligand-activated VDR competed with T cell transcription factor (TCF)-4 for beta-catenin binding. Accordingly, 1alpha,25(OH)2D(3) repressed beta-catenin-TCF-4 transcriptional activity. Moreover, VDR activity was enhanced by ectopic beta-catenin and reduced by TCF-4. Also, 1alpha,25(OH)2D(3) inhibited expression of beta-catenin-TCF-4-responsive genes, c-myc, peroxisome proliferator-activated receptor delta, Tcf-1, and CD44, whereas it induced expression of ZO-1. Our results show that 1alpha,25(OH)2D(3) induces E-cadherin and modulates beta-catenin-TCF-4 target genes in a manner opposite to that of beta-catenin, promoting the differentiation of colon carcinoma cells.

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Induction of colocalization of E-cadherin and β-catenin at the plasma membrane by 1α,25(OH)2D3. Analysis by immunofluorescence and confocal laser scanning microscopy of the expression of these two proteins in SW480-ADH or SW480-R cells at 48 h after either treatment with 1α,25(OH)2D3 (10−7 M) or transfection with an expression vector for human E-cadherin. Double immunofluorescence was performed using anti–E-cadherin and anti–β-catenin antibodies followed by the addition of the corresponding secondary TRICT-conjugated (E-cadherin, red) or FITC-conjugated (β-catenin, green) antibodies. The merge of both signals (yellow) indicates the areas of colocalization of both proteins. Bars, 10 μm.
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fig5: Induction of colocalization of E-cadherin and β-catenin at the plasma membrane by 1α,25(OH)2D3. Analysis by immunofluorescence and confocal laser scanning microscopy of the expression of these two proteins in SW480-ADH or SW480-R cells at 48 h after either treatment with 1α,25(OH)2D3 (10−7 M) or transfection with an expression vector for human E-cadherin. Double immunofluorescence was performed using anti–E-cadherin and anti–β-catenin antibodies followed by the addition of the corresponding secondary TRICT-conjugated (E-cadherin, red) or FITC-conjugated (β-catenin, green) antibodies. The merge of both signals (yellow) indicates the areas of colocalization of both proteins. Bars, 10 μm.

Mentions: Exogenous E-cadherin induces β-catenin relocation by nuclear export and the formation of E-cadherin–β-catenin complexes at the plasma membrane in SW480 and other cell lines lacking endogenous E-cadherin (Sadot et al., 1998; Orsulic et al., 1999). Since 1α,25(OH)2D3 induced E-cadherin expression in SW480-ADH cells, we determined whether it could promote a similar effect in these cells. 2-d treatments with 1α,25(OH)2D3 induced a nearly complete colocalization of both proteins at the region of the plasma membrane in contact with neighboring cells (Fig. 5 , top). An identical effect was observed when untreated SW480-ADH cells were transfected with an E-cadherin expression plasmid (Fig. 5, top). As controls, we used VDR-negative SW480-R cells, which did not show any change in the strong nuclear β-catenin staining upon 1α,25(OH)2D3 addition (Fig. 5, bottom). As expected, exogenous E-cadherin caused the same change in β-catenin localization as in SW480-ADH cells (Fig. 5, bottom). To correlate the change in β-catenin localization induced by 1α,25(OH)2D3 with the effects on β-catenin–TCF/LEF-1 target genes as a function of time, we studied the kinetics of β-catenin nuclear export. To this end, we analyzed the immunofluorescence of SW480-ADH cells stained with an anti–β-catenin antibody by confocal laser microscopy and counted the cells displaying predominantly nuclear, a mixed nuclear-cytosolic, or an exclusively membranous β-catenin localization (Fig. 6 A, insets). The number of nuclear-positive β-catenin cells clearly decreased (from 66 to 24%) as soon as 16 h after 1α,25(OH)2D3 addition, was further reduced (to 10%) at 48 h, and was extremely low (around 2%) 7 d later (Fig. 6 A, white bars). Conversely, the proportion of cells with predominant membrane-bound β-catenin increased progressively from 11% in nontreated cultures to 70% after 7 d of 1α,25(OH)2D3 treatment (Fig. 6 A, black bars).


Vitamin D(3) promotes the differentiation of colon carcinoma cells by the induction of E-cadherin and the inhibition of beta-catenin signaling.

Pálmer HG, González-Sancho JM, Espada J, Berciano MT, Puig I, Baulida J, Quintanilla M, Cano A, de Herreros AG, Lafarga M, Muñoz A - J. Cell Biol. (2001)

Induction of colocalization of E-cadherin and β-catenin at the plasma membrane by 1α,25(OH)2D3. Analysis by immunofluorescence and confocal laser scanning microscopy of the expression of these two proteins in SW480-ADH or SW480-R cells at 48 h after either treatment with 1α,25(OH)2D3 (10−7 M) or transfection with an expression vector for human E-cadherin. Double immunofluorescence was performed using anti–E-cadherin and anti–β-catenin antibodies followed by the addition of the corresponding secondary TRICT-conjugated (E-cadherin, red) or FITC-conjugated (β-catenin, green) antibodies. The merge of both signals (yellow) indicates the areas of colocalization of both proteins. Bars, 10 μm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2150773&req=5

fig5: Induction of colocalization of E-cadherin and β-catenin at the plasma membrane by 1α,25(OH)2D3. Analysis by immunofluorescence and confocal laser scanning microscopy of the expression of these two proteins in SW480-ADH or SW480-R cells at 48 h after either treatment with 1α,25(OH)2D3 (10−7 M) or transfection with an expression vector for human E-cadherin. Double immunofluorescence was performed using anti–E-cadherin and anti–β-catenin antibodies followed by the addition of the corresponding secondary TRICT-conjugated (E-cadherin, red) or FITC-conjugated (β-catenin, green) antibodies. The merge of both signals (yellow) indicates the areas of colocalization of both proteins. Bars, 10 μm.
Mentions: Exogenous E-cadherin induces β-catenin relocation by nuclear export and the formation of E-cadherin–β-catenin complexes at the plasma membrane in SW480 and other cell lines lacking endogenous E-cadherin (Sadot et al., 1998; Orsulic et al., 1999). Since 1α,25(OH)2D3 induced E-cadherin expression in SW480-ADH cells, we determined whether it could promote a similar effect in these cells. 2-d treatments with 1α,25(OH)2D3 induced a nearly complete colocalization of both proteins at the region of the plasma membrane in contact with neighboring cells (Fig. 5 , top). An identical effect was observed when untreated SW480-ADH cells were transfected with an E-cadherin expression plasmid (Fig. 5, top). As controls, we used VDR-negative SW480-R cells, which did not show any change in the strong nuclear β-catenin staining upon 1α,25(OH)2D3 addition (Fig. 5, bottom). As expected, exogenous E-cadherin caused the same change in β-catenin localization as in SW480-ADH cells (Fig. 5, bottom). To correlate the change in β-catenin localization induced by 1α,25(OH)2D3 with the effects on β-catenin–TCF/LEF-1 target genes as a function of time, we studied the kinetics of β-catenin nuclear export. To this end, we analyzed the immunofluorescence of SW480-ADH cells stained with an anti–β-catenin antibody by confocal laser microscopy and counted the cells displaying predominantly nuclear, a mixed nuclear-cytosolic, or an exclusively membranous β-catenin localization (Fig. 6 A, insets). The number of nuclear-positive β-catenin cells clearly decreased (from 66 to 24%) as soon as 16 h after 1α,25(OH)2D3 addition, was further reduced (to 10%) at 48 h, and was extremely low (around 2%) 7 d later (Fig. 6 A, white bars). Conversely, the proportion of cells with predominant membrane-bound β-catenin increased progressively from 11% in nontreated cultures to 70% after 7 d of 1α,25(OH)2D3 treatment (Fig. 6 A, black bars).

Bottom Line: Accordingly, 1alpha,25(OH)2D(3) repressed beta-catenin-TCF-4 transcriptional activity.Also, 1alpha,25(OH)2D(3) inhibited expression of beta-catenin-TCF-4-responsive genes, c-myc, peroxisome proliferator-activated receptor delta, Tcf-1, and CD44, whereas it induced expression of ZO-1.Our results show that 1alpha,25(OH)2D(3) induces E-cadherin and modulates beta-catenin-TCF-4 target genes in a manner opposite to that of beta-catenin, promoting the differentiation of colon carcinoma cells.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Investigaciones Biomédicas "Alberto Sols, " Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, E-28029 Madrid, Spain.

ABSTRACT
The beta-catenin signaling pathway is deregulated in nearly all colon cancers. Nonhypercalcemic vitamin D3 (1alpha,25-dehydroxyvitamin D(3)) analogues are candidate drugs to treat this neoplasia. We show that these compounds promote the differentiation of human colon carcinoma SW480 cells expressing vitamin D receptors (VDRs) (SW480-ADH) but not that of a malignant subline (SW480-R) or metastasic derivative (SW620) cells lacking VDR. 1alpha,25(OH)2D(3) induced the expression of E-cadherin and other adhesion proteins (occludin, Zonula occludens [ZO]-1, ZO-2, vinculin) and promoted the translocation of beta-catenin, plakoglobin, and ZO-1 from the nucleus to the plasma membrane. Ligand-activated VDR competed with T cell transcription factor (TCF)-4 for beta-catenin binding. Accordingly, 1alpha,25(OH)2D(3) repressed beta-catenin-TCF-4 transcriptional activity. Moreover, VDR activity was enhanced by ectopic beta-catenin and reduced by TCF-4. Also, 1alpha,25(OH)2D(3) inhibited expression of beta-catenin-TCF-4-responsive genes, c-myc, peroxisome proliferator-activated receptor delta, Tcf-1, and CD44, whereas it induced expression of ZO-1. Our results show that 1alpha,25(OH)2D(3) induces E-cadherin and modulates beta-catenin-TCF-4 target genes in a manner opposite to that of beta-catenin, promoting the differentiation of colon carcinoma cells.

Show MeSH
Related in: MedlinePlus