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Article

Serotonin inhibits the influx of iron into cells by decreasing the expression of SLC11A2 in Caco‑2 cells

  • Authors:
    • Yuta Sugino
    • Momoko Ameku
    • Norika Kawanishi
    • Shinpei Nonen
  • View Affiliations / Copyright

    Affiliations: School of Pharmacy, Hyogo Medical University, Kobe, Hyogo 650‑8530, Japan
  • Article Number: 13
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    Published online on: November 14, 2025
       https://doi.org/10.3892/br.2025.2086
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Abstract

Iron is an essential trace element for biological functions and its homeostasis is strictly regulated. Intestinal iron absorption primarily occurs in the duodenum, and the iron transporter solute carrier family 11 member 2 (SLC11A2; also known as divalent metal transporter 1) is crucial in this process. Notably, iron absorption is regulated by iron stores in the body and hormones; however, the effects of serotonin on iron transport are poorly understood. The present study aimed to analyze the effects of serotonin on the expression and function of SLC11A2 using human colon carcinoma‑derived Caco‑2 cells. SLC11A2 mRNA and protein levels were measured using reverse transcription‑quantitative PCR and western blotting, respectively. To investigate the effect of serotonin on intracellular iron concentrations, iron levels were measured in cells treated with a soluble iron salt. The results revealed that serotonin decreased SLC11A2 mRNA levels to ~25% and protein levels to ~35% of control values (P<0.01). A soluble iron salt markedly increased the intracellular iron concentration in Caco‑2 cells; however, this increase was suppressed by serotonin treatment (P<0.01). To investigate the mechanism of serotonin action, Caco‑2 cells were treated with BW723C86, a selective 5‑hydroxytryptamine (5‑HT) 2B receptor agonist. BW723C86 also decreased SLC11A2 mRNA levels to ~25% and protein levels to ~60% of control values (P<0.01 and P<0.05). Furthermore, BW723C86 suppressed the increase in intracellular iron concentration induced by a soluble iron salt (P<0.01). In addition, the serotonin‑induced decrease in SLC11A2 mRNA levels was inhibited by SB204741, a selective antagonist of the 5‑HT2B receptor (P<0.05). These findings suggested a novel link between serotonin signaling and iron absorption, which may inform future therapeutic approaches for iron deficiency anemia.
View Figures

Figure 1

Effect of serotonin on the mRNA
expression of SLC11A2 in Caco-2 cells. (A) Cells were treated with
5-HT at the indicated concentrations for 6 h. (B) Cells were
treated with 500 µg/ml 5-HT for the indicated times. The mRNA
levels of SLC11A2 were measured using reverse
transcription-quantitative PCR and normalized to that of GAPDH.
Results show the mean ± standard error (n=4). Statistical
significance was determined using one-way ANOVA followed by the
Tukey-Kramer post hoc test. *P<0.05;
**P<0.01 vs. control. Exact P-values: (A) 0.014,
<0.001, <0.001, <0.001; (B) 0.013, <0.001, <0.001,
<0.001, <0.001. SLC11A2, solute carrier family 11 member 2;
5-HT, serotonin.

Figure 2

Effect of serotonin on the protein
expression of SLC11A2 and the intracellular iron-ion concentration
in Caco-2 cells. (A) Cells were treated with 500 µg/ml 5-HT for the
indicated times. Western blotting was conducted to detect SLC11A2
protein. Panels show the western blotting results indicating the
SLC11A2 expression of cells. Bar graph indicates the relative
intensity of SLC11A2 signals shown at panels, quantified with a
densitometer. Result shows the mean ± SE (n=4). Statistical
significance was determined by one-way ANOVA followed by the
Tukey-Kramer post hoc test. **P<0.01 vs. control.
Exact P-values: <0.001, <0.001. (B) Cells were treated with
500 µg/ml 5-HT for 6 h or 24 h and then exposed to 10 mM ammonium
iron (II) sulfate hexahydrate for 30 min. Intracellular iron
concentration was determined by ferrozine assay. Result shows the
mean ± standard error (n=4). Statistical significance was
determined using one-way ANOVA followed by the Tukey-Kramer post
hoc test. **P<0.01. Exact P-values: <0.001,
<0.001. SLC11A2, solute carrier family 11 member 2; 5-HT,
serotonin.

Figure 3

mRNA expression of serotonin receptor
subunits in Caco-2 cells. The mRNA expression of each serotonin
receptor in the cells was examined by reverse trasnscription-PCR.
(A) Agarose gel electrophoresis of the PCR amplified products of
GAPDH (lane G) and serotonin receptor subtypes (lane 1-10). Lane M
represents a 100-bp DNA ladder. (B) The relative intensity of the
bands was quantified by densitometry using ImageJ software, and the
mRNA levels were normalized to GAPDH. Data are expressed relative
to the maximum expression level, which was arbitrarily set to 1.0.
Results are representative of four independent experiments. 5-HT,
serotonin.

Figure 4

Effect of BW on the mRNA and protein
expression of SLC11A2 and the intracellular iron-ion concentration
in Caco-2 cells. (A) Cells were treated with 100 µM BW for the
indicated times. The mRNA levels of SLC11A2 were measured by
reverse transcription-quantitative PCR and normalized to that of
GAPDH. Results show the mean ± SE (n=4). Statistical significance
was determined using one-way ANOVA followed by the Tukey-Kramer
post hoc test. **P<0.01 vs. control. Exact P-values:
<0.001, <0.001, <0.001. (B) Cells were treated with 100 µM
BW for the indicated times. Western blotting was conducted to
detect SLC11A2 protein. Panels show the western blotting results
indicating the SLC11A2 expression of cells. Bar graph indicates the
relative intensity of SLC11A2 signals shown at panels, quantified
with a densitometer. Result shows the mean ± SE (n=4). Statistical
significance was determined by one-way ANOVA followed by the
Tukey-Kramer post hoc test (*P<0.05 vs. control).
Exact P-values: 0.039, 0.043. (C) Cells were treated with 100 µM BW
for 6 h or 24 h and then exposed to 10 mM ammonium iron (II)
sulfate hexahydrate for 30 min. Intracellular iron concentration
was determined by ferrozine assay. Result shows the mean ± SE
(n=4). Statistical significance was determined using one-way ANOVA
followed by the Tukey-Kramer post hoc test. **P<0.01.
Exact P-values: <0.001, 0.009. BW, BW723C86; SLC11A2, solute
carrier family 11 member 2; SE, standard error.

Figure 5

Effect of SB204741 on the
serotonin-induced reduction in SLC11A2 mRNA expression in Caco-2
cells. Cells were treated with SB at the indicated concentrations
for 24 h, followed by 500 µg/ml 5-HT for 6 h. The mRNA levels of
SLC11A2 were measured using reverse transcription-quantitative PCR
and normalized to that of GAPDH. Results show the mean ± standard
error (n=4). Statistical significance was determined using one-way
ANOVA followed by the Tukey-Kramer post hoc test.
*P<0.05; **P<0.01. Exact P-values:
<0.001, 0.036. SLC11A2, solute carrier family 11 member 2; 5-HT,
serotonin; SB, SB204741.
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Copy and paste a formatted citation
Spandidos Publications style
Sugino Y, Ameku M, Kawanishi N and Nonen S: Serotonin inhibits the influx of iron into cells by decreasing the expression of SLC11A2 in Caco‑2 cells. Biomed Rep 24: 13, 2026.
APA
Sugino, Y., Ameku, M., Kawanishi, N., & Nonen, S. (2026). Serotonin inhibits the influx of iron into cells by decreasing the expression of SLC11A2 in Caco‑2 cells. Biomedical Reports, 24, 13. https://doi.org/10.3892/br.2025.2086
MLA
Sugino, Y., Ameku, M., Kawanishi, N., Nonen, S."Serotonin inhibits the influx of iron into cells by decreasing the expression of SLC11A2 in Caco‑2 cells". Biomedical Reports 24.1 (2026): 13.
Chicago
Sugino, Y., Ameku, M., Kawanishi, N., Nonen, S."Serotonin inhibits the influx of iron into cells by decreasing the expression of SLC11A2 in Caco‑2 cells". Biomedical Reports 24, no. 1 (2026): 13. https://doi.org/10.3892/br.2025.2086
Copy and paste a formatted citation
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Spandidos Publications style
Sugino Y, Ameku M, Kawanishi N and Nonen S: Serotonin inhibits the influx of iron into cells by decreasing the expression of SLC11A2 in Caco‑2 cells. Biomed Rep 24: 13, 2026.
APA
Sugino, Y., Ameku, M., Kawanishi, N., & Nonen, S. (2026). Serotonin inhibits the influx of iron into cells by decreasing the expression of SLC11A2 in Caco‑2 cells. Biomedical Reports, 24, 13. https://doi.org/10.3892/br.2025.2086
MLA
Sugino, Y., Ameku, M., Kawanishi, N., Nonen, S."Serotonin inhibits the influx of iron into cells by decreasing the expression of SLC11A2 in Caco‑2 cells". Biomedical Reports 24.1 (2026): 13.
Chicago
Sugino, Y., Ameku, M., Kawanishi, N., Nonen, S."Serotonin inhibits the influx of iron into cells by decreasing the expression of SLC11A2 in Caco‑2 cells". Biomedical Reports 24, no. 1 (2026): 13. https://doi.org/10.3892/br.2025.2086
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