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Selective alteration of gene expression in response to natural and synthetic retinoids.

Brand C, Ségard P, Plouvier P, Formstecher P, Danzé PM, Lefebvre P - BMC Pharmacol. (2002)

Bottom Line: This suggested that gene-selective modulation could be achieved by structurally distinct retinoids.Furthermore, this differential ability to regulate promoter activities was also observed in murine P19 cells for the RARbeta2 and CRABPII gene, showing conclusively that retinoid structure has a dramatic impact on the regulation of endogenous genes.Our findings therefore show that some degree of selective induction or repression of gene expression may be achieved when using appropriately designed ligands for retinoic acid receptors, extending the concept of selective modulators from estrogen and peroxisome proliferator activated receptors to the class of retinoid receptors.

View Article: PubMed Central - HTML - PubMed

Affiliation: INSERM U 459 and Ligue nationale contre le Cancer, Faculté de Médecine Henri Warembourg, 1, place de Verdun, 59045 Lille cedex, France. brand@lille.inserm.fr

ABSTRACT

Background: Retinoids are very potent inducers of cellular differentiation and apoptosis, and are efficient anti-tumoral agents. Synthetic retinoids are designed to restrict their toxicity and side effects, mostly by increasing their selectivity toward each isotype of retinoic acids receptors (RARalpha,beta, gamma and RXRalpha, beta, gamma). We however previously showed that retinoids displayed very different abilities to activate retinoid-inducible reporter genes, and that these differential properties were correlated to the ability of a given ligand to promote SRC-1 recruitment by DNA-bound RXR:RAR heterodimers. This suggested that gene-selective modulation could be achieved by structurally distinct retinoids.

Results: Using the differential display mRNA technique, we identified several genes on the basis of their differential induction by natural or synthetic retinoids in human cervix adenocarcinoma cells. Furthermore, this differential ability to regulate promoter activities was also observed in murine P19 cells for the RARbeta2 and CRABPII gene, showing conclusively that retinoid structure has a dramatic impact on the regulation of endogenous genes.

Conclusions: Our findings therefore show that some degree of selective induction or repression of gene expression may be achieved when using appropriately designed ligands for retinoic acid receptors, extending the concept of selective modulators from estrogen and peroxisome proliferator activated receptors to the class of retinoid receptors.

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Related in: MedlinePlus

Expression of TAXREB107 mRNA in HeLa cells treated with different ligands of the retinoic acid receptor. HeLa cells were treated with the different ligands for 4 hours. Total RNA (20 μg) was probed sequentially with fluorescein-labeled partial human TAXREB cDNA and 18S rRNA probes. Blots were quantified using a Storm™ apparatus. Values for the TAXREB transcript were normalized to the 18S rRNA level. A) Upper panel: TAXREB107 transcript in HeLa cells treated with various retinoids. Lower panel: 18S rRNA. B) Quantification of TAXREB107 expression. Results are presented as the mean +/- S.E.M. of three different experiments. Retinoid concentrations were as in Figure 3.
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Figure 4: Expression of TAXREB107 mRNA in HeLa cells treated with different ligands of the retinoic acid receptor. HeLa cells were treated with the different ligands for 4 hours. Total RNA (20 μg) was probed sequentially with fluorescein-labeled partial human TAXREB cDNA and 18S rRNA probes. Blots were quantified using a Storm™ apparatus. Values for the TAXREB transcript were normalized to the 18S rRNA level. A) Upper panel: TAXREB107 transcript in HeLa cells treated with various retinoids. Lower panel: 18S rRNA. B) Quantification of TAXREB107 expression. Results are presented as the mean +/- S.E.M. of three different experiments. Retinoid concentrations were as in Figure 3.

Mentions: Search against EMBL/GenBank databases using the Blast server identified the human initiation factor 4B (hIF4B, SRIG 69), human apoferritin H (SRIG 157) and human TAXREB 107 (SRIG 177). We also noted that the human plasminogen activator was identified in this screen, which is a gene known to be activated by retinoids [33], as well as the ubiquitin conjugating enzyme UCE, which was identified recently by a similar approach as an atRA-inducible gene in acute promyelocytic leukemia (APL) cells [33,34]. Northern blot analysis of HeLa cells RNA was thus used to assay the rate of expression of several genes after retinoid treatment and results are shown for ferritin H and TAXREB 107 (Figure 3 and 4).


Selective alteration of gene expression in response to natural and synthetic retinoids.

Brand C, Ségard P, Plouvier P, Formstecher P, Danzé PM, Lefebvre P - BMC Pharmacol. (2002)

Expression of TAXREB107 mRNA in HeLa cells treated with different ligands of the retinoic acid receptor. HeLa cells were treated with the different ligands for 4 hours. Total RNA (20 μg) was probed sequentially with fluorescein-labeled partial human TAXREB cDNA and 18S rRNA probes. Blots were quantified using a Storm™ apparatus. Values for the TAXREB transcript were normalized to the 18S rRNA level. A) Upper panel: TAXREB107 transcript in HeLa cells treated with various retinoids. Lower panel: 18S rRNA. B) Quantification of TAXREB107 expression. Results are presented as the mean +/- S.E.M. of three different experiments. Retinoid concentrations were as in Figure 3.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC113761&req=5

Figure 4: Expression of TAXREB107 mRNA in HeLa cells treated with different ligands of the retinoic acid receptor. HeLa cells were treated with the different ligands for 4 hours. Total RNA (20 μg) was probed sequentially with fluorescein-labeled partial human TAXREB cDNA and 18S rRNA probes. Blots were quantified using a Storm™ apparatus. Values for the TAXREB transcript were normalized to the 18S rRNA level. A) Upper panel: TAXREB107 transcript in HeLa cells treated with various retinoids. Lower panel: 18S rRNA. B) Quantification of TAXREB107 expression. Results are presented as the mean +/- S.E.M. of three different experiments. Retinoid concentrations were as in Figure 3.
Mentions: Search against EMBL/GenBank databases using the Blast server identified the human initiation factor 4B (hIF4B, SRIG 69), human apoferritin H (SRIG 157) and human TAXREB 107 (SRIG 177). We also noted that the human plasminogen activator was identified in this screen, which is a gene known to be activated by retinoids [33], as well as the ubiquitin conjugating enzyme UCE, which was identified recently by a similar approach as an atRA-inducible gene in acute promyelocytic leukemia (APL) cells [33,34]. Northern blot analysis of HeLa cells RNA was thus used to assay the rate of expression of several genes after retinoid treatment and results are shown for ferritin H and TAXREB 107 (Figure 3 and 4).

Bottom Line: This suggested that gene-selective modulation could be achieved by structurally distinct retinoids.Furthermore, this differential ability to regulate promoter activities was also observed in murine P19 cells for the RARbeta2 and CRABPII gene, showing conclusively that retinoid structure has a dramatic impact on the regulation of endogenous genes.Our findings therefore show that some degree of selective induction or repression of gene expression may be achieved when using appropriately designed ligands for retinoic acid receptors, extending the concept of selective modulators from estrogen and peroxisome proliferator activated receptors to the class of retinoid receptors.

View Article: PubMed Central - HTML - PubMed

Affiliation: INSERM U 459 and Ligue nationale contre le Cancer, Faculté de Médecine Henri Warembourg, 1, place de Verdun, 59045 Lille cedex, France. brand@lille.inserm.fr

ABSTRACT

Background: Retinoids are very potent inducers of cellular differentiation and apoptosis, and are efficient anti-tumoral agents. Synthetic retinoids are designed to restrict their toxicity and side effects, mostly by increasing their selectivity toward each isotype of retinoic acids receptors (RARalpha,beta, gamma and RXRalpha, beta, gamma). We however previously showed that retinoids displayed very different abilities to activate retinoid-inducible reporter genes, and that these differential properties were correlated to the ability of a given ligand to promote SRC-1 recruitment by DNA-bound RXR:RAR heterodimers. This suggested that gene-selective modulation could be achieved by structurally distinct retinoids.

Results: Using the differential display mRNA technique, we identified several genes on the basis of their differential induction by natural or synthetic retinoids in human cervix adenocarcinoma cells. Furthermore, this differential ability to regulate promoter activities was also observed in murine P19 cells for the RARbeta2 and CRABPII gene, showing conclusively that retinoid structure has a dramatic impact on the regulation of endogenous genes.

Conclusions: Our findings therefore show that some degree of selective induction or repression of gene expression may be achieved when using appropriately designed ligands for retinoic acid receptors, extending the concept of selective modulators from estrogen and peroxisome proliferator activated receptors to the class of retinoid receptors.

Show MeSH
Related in: MedlinePlus