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All-Trans Retinoic Acid Increases Aquaporin 3 Expression in Human Vaginal Epithelial Cells

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Water channel aquaporin 3 (AQP3) is an aquaglyceroporin that transports small neutral solutes and water. All-trans retinoic acid (ATRA), a member of the retinoid drug class, acts as a regulator in several biological processes.

Aim: To investigate the effect of ATRA on the expression of AQP3 in human vaginal epithelial cells.

Methods: Human vaginal mucosal epithelial cells (CRL2616) were treated with ATRA 0, 0.01, 0.1, and 1 μmol/L for 24 hours to examine the dose-dependent effects of ATRA and with ATRA 1 μmol/L for 0, 3, 6, 12, and 24 hours.

Main outcome measures: The expression of AQP3 and retinoic acid receptor (RAR) was determined by western blot analysis and reverse transcription polymerase chain reaction.

Results: AQP3 was detected in the cell membrane of human vaginal epithelial cells. ATRA increased the protein expression and mRNA levels of AQP3 in a dose-dependent manner (P < .05). ATRA also increased the protein expression of RARα (P < .05). Treatment of CRL2616 cells with an RAR antagonist (Ro 41-5253) significantly decreased AQP3 protein expression (P < .05).

Conclusion: ATRA mediated by RARα increased AQP3 gene and protein expression in human vaginal mucosal epithelial cells. These results imply that AQP3 regulated by ATRA could play an important role in the mechanism of vaginal lubrication.

No MeSH data available.


Effect of all-trans retinoic acid on the expression of AQP3 protein in cultured human vaginal mucosal epithelial cells. Panel A shows the detection of AQP3 in the cell membrane of human vaginal epithelial cells (green) by confocal microscopy after immunostaining with anti-AQP3. Cells also were stained with 4′-6-diamidino-2-phenylindole to visualize the nucleus (blue). Panel B shows the incubation of cells with all-trans retinoic acid 1 μmol/L for 0, 3, 6, 12, or 24 hours. Twenty micrograms of whole-cell lysates was applied to sodium dodecylsulfate polyacrylamide gel electrophoresis followed by western blotting with anti-AQP3 and anti–β-actin antibodies. Panel C shows the incubation of cells with all-trans retinoic acid 0, 0.01, 0.1, or 1 μmol/L for 24 hours. Levels of AQP3 protein were examined by western blotting. The results are representative of three independent experiments. The expression of AQP3 was dose-dependently increased by all-trans retinoic acid treatment. ∗P < 0.05 vs untreated control. AQP3 = aquaporin 3; Con = control; GAPDH = glyceraldehyde 3-phosphate dehydrogenase.
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fig1: Effect of all-trans retinoic acid on the expression of AQP3 protein in cultured human vaginal mucosal epithelial cells. Panel A shows the detection of AQP3 in the cell membrane of human vaginal epithelial cells (green) by confocal microscopy after immunostaining with anti-AQP3. Cells also were stained with 4′-6-diamidino-2-phenylindole to visualize the nucleus (blue). Panel B shows the incubation of cells with all-trans retinoic acid 1 μmol/L for 0, 3, 6, 12, or 24 hours. Twenty micrograms of whole-cell lysates was applied to sodium dodecylsulfate polyacrylamide gel electrophoresis followed by western blotting with anti-AQP3 and anti–β-actin antibodies. Panel C shows the incubation of cells with all-trans retinoic acid 0, 0.01, 0.1, or 1 μmol/L for 24 hours. Levels of AQP3 protein were examined by western blotting. The results are representative of three independent experiments. The expression of AQP3 was dose-dependently increased by all-trans retinoic acid treatment. ∗P < 0.05 vs untreated control. AQP3 = aquaporin 3; Con = control; GAPDH = glyceraldehyde 3-phosphate dehydrogenase.

Mentions: AQP3 was expressed in the cell membrane of human vaginal mucosal epithelial cells (Figure 1A).


All-Trans Retinoic Acid Increases Aquaporin 3 Expression in Human Vaginal Epithelial Cells
Effect of all-trans retinoic acid on the expression of AQP3 protein in cultured human vaginal mucosal epithelial cells. Panel A shows the detection of AQP3 in the cell membrane of human vaginal epithelial cells (green) by confocal microscopy after immunostaining with anti-AQP3. Cells also were stained with 4′-6-diamidino-2-phenylindole to visualize the nucleus (blue). Panel B shows the incubation of cells with all-trans retinoic acid 1 μmol/L for 0, 3, 6, 12, or 24 hours. Twenty micrograms of whole-cell lysates was applied to sodium dodecylsulfate polyacrylamide gel electrophoresis followed by western blotting with anti-AQP3 and anti–β-actin antibodies. Panel C shows the incubation of cells with all-trans retinoic acid 0, 0.01, 0.1, or 1 μmol/L for 24 hours. Levels of AQP3 protein were examined by western blotting. The results are representative of three independent experiments. The expression of AQP3 was dose-dependently increased by all-trans retinoic acid treatment. ∗P < 0.05 vs untreated control. AQP3 = aquaporin 3; Con = control; GAPDH = glyceraldehyde 3-phosphate dehydrogenase.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5121539&req=5

fig1: Effect of all-trans retinoic acid on the expression of AQP3 protein in cultured human vaginal mucosal epithelial cells. Panel A shows the detection of AQP3 in the cell membrane of human vaginal epithelial cells (green) by confocal microscopy after immunostaining with anti-AQP3. Cells also were stained with 4′-6-diamidino-2-phenylindole to visualize the nucleus (blue). Panel B shows the incubation of cells with all-trans retinoic acid 1 μmol/L for 0, 3, 6, 12, or 24 hours. Twenty micrograms of whole-cell lysates was applied to sodium dodecylsulfate polyacrylamide gel electrophoresis followed by western blotting with anti-AQP3 and anti–β-actin antibodies. Panel C shows the incubation of cells with all-trans retinoic acid 0, 0.01, 0.1, or 1 μmol/L for 24 hours. Levels of AQP3 protein were examined by western blotting. The results are representative of three independent experiments. The expression of AQP3 was dose-dependently increased by all-trans retinoic acid treatment. ∗P < 0.05 vs untreated control. AQP3 = aquaporin 3; Con = control; GAPDH = glyceraldehyde 3-phosphate dehydrogenase.
Mentions: AQP3 was expressed in the cell membrane of human vaginal mucosal epithelial cells (Figure 1A).

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Water channel aquaporin 3 (AQP3) is an aquaglyceroporin that transports small neutral solutes and water. All-trans retinoic acid (ATRA), a member of the retinoid drug class, acts as a regulator in several biological processes.

Aim: To investigate the effect of ATRA on the expression of AQP3 in human vaginal epithelial cells.

Methods: Human vaginal mucosal epithelial cells (CRL2616) were treated with ATRA 0, 0.01, 0.1, and 1 &mu;mol/L for 24 hours to examine the dose-dependent effects of ATRA and with ATRA 1 &mu;mol/L for 0, 3, 6, 12, and 24 hours.

Main outcome measures: The expression of AQP3 and retinoic acid receptor (RAR) was determined by western blot analysis and reverse transcription polymerase chain reaction.

Results: AQP3 was detected in the cell membrane of human vaginal epithelial cells. ATRA increased the protein expression and mRNA levels of AQP3 in a dose-dependent manner (P &lt; .05). ATRA also increased the protein expression of RAR&alpha; (P &lt; .05). Treatment of CRL2616 cells with an RAR antagonist (Ro 41-5253) significantly decreased AQP3 protein expression (P &lt; .05).

Conclusion: ATRA mediated by RAR&alpha; increased AQP3 gene and protein expression in human vaginal mucosal epithelial cells. These results imply that AQP3 regulated by ATRA could play an important role in the mechanism of vaginal lubrication.

No MeSH data available.