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Article Open Access

ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis

  • Authors:
    • Yin Tian
    • Yun Liu
    • Yangyang Shang
    • Lijian Ran
    • Li Liu
    • Rongquan Wang
    • Jun Ye
  • View Affiliations / Copyright

    Affiliations: Department of Gastroenterology, The People's Hospital of Yubei District of Chongqing City, Chongqing 401120, P.R. China, Institute of Gastroenterology of People's Liberation Army, Southwest Hospital, Army Medical University, Chongqing 400038, P.R. China
    Copyright: © Tian et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 73
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    Published online on: December 18, 2025
       https://doi.org/10.3892/mmr.2025.13783
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Abstract

The pathogenesis of inflammatory bowel disease is associated with dysfunction of the intestinal mucosal barrier. Protein sialylation serves an important role in maintaining the integrity of this barrier. The present study investigated how α2,3‑linked sialylation catalyzed by protein ST3Gal1 affected intestinal barrier function and impacted the pathogenesis of human ulcerative colitis (UC). The present study employed Caco‑2, HT29‑MTX‑E12 and THP‑1 cells with distinct functionalities to establish an in vitro triple‑culture model. This model was utilized to simulate both healthy and inflamed states of the human intestine for investigating the impact of ST3Gal1‑mediated α2,3‑sialylation on the integrity of the intestinal barrier. The triple‑culture model was stably infected with adenoviral particles or lentiviral vectors to establish ST3Gal1 knockdown and overexpression, respectively, followed by isolation through incubation with 4 µg/ml puromycin. The functionality of the intestinal barrier was assessed via trans‑epithelial electrical resistance and FITC‑dextran permeability assays. ST3Gal1 expression was found to be associated with inflammation of the intestinal mucosa in patients with UC and a mouse model of dextran sulfate sodium‑induced colitis. Notably, suppressed expression of ST3Gal1 in the intestinal epithelial cell (IEC) monolayer enhanced the functionality of the intestinal barrier, whereas its overexpression caused intestinal barrier function deterioration. ST3Gal1 expression in the IEC monolayer altered the expression of intestinal mucus barrier‑associated mucin 2 (MUC2) and trefoil factor 3 (TFF3), goblet cell differentiation‑associated homeobox protein CDX‑2 (CDX2), inflammation‑associated phosphorylated (p)‑STAT3, and the inflammatory mediators IL‑1β, IL‑6 and IL‑8. Specifically, MUC2, TFF3 and CDX2 were positively associated with enhanced barrier integrity, whereas p‑STAT3, IL‑1β, IL‑6 and IL‑8 were negatively correlated with barrier function. Collectively, these results demonstrated a strong association between these factors and the regulation of intestinal barrier function. In conclusion, ST3Gal1‑catalyzed α2,3‑linkage formation in IECs may be closely associated with intestinal barrier function via its effect on the expression of barrier‑associated proteins and inflammatory mediators related to intestinal mucosa inflammation. 
View Figures

Figure 1

ST3Gal1 mRNA levels in intestinal
mucosa from patients with UC and a mouse DSS-induced colitis model
are associated with inflammatory states. (A) Comparison of ST3Gal1
mRNA levels between human normal colon mucosa from healthy controls
(n=5) and colonic mucosa from patients diagnosed with UC (n=21) in
the GDS3119 dataset from the GEO database. (B) ST3Gal1 mRNA levels
in human normal colonic tissues from healthy controls (n=5),
inflamed colonic tissues from patients with UC and visible
macroscopic inflammation (n=8), and non-inflamed colonic mucosal
tissues from patients with UC without macroscopic signs of
inflammation (n=13) in the GEO dataset GDS3119. (C) ST3Gal1 mRNA
levels in non-lesional mucosa (n=44) and lesional mucosa (n=75)
from different patients with UC, obtained from the GEO dataset
GSE107499. (D) ST3Gal1 mRNA levels in the normal mucosa (n=21) from
healthy controls and the inflamed mucosa (n=162) from patients with
UC with varying modified Mayo scores in the GEO dataset GSE92415.
(E) ST3Gal1 mRNA levels in colon tissues from mice treated with DSS
to induce colitis (independent, non-overlapping mice) divided into
four groups: Before DSS administration (n=5), and at 2, 4 and 6
days post-DSS treatment (n=6/group). Data were obtained from the
GEO dataset GDS3859. Data in A, C and D are presented as box and
whisker plots, whereas data in B and E are presented as the mean ±
SD. Statistical significance was determined using specific tests
for each subpart based on group comparisons: (A and C) Unpaired
t-test and (B, D and E) one-way analysis of variance followed by
Tukey's post-hoc test. *P<0.05 and **P<0.01. DSS, dextran
sulfate sodium; GEO, Gene Expression Omnibus; UC, ulcerative
colitis.

Figure 2

Establishment of ST3Gal1-interfered
IEC and ST3Gal1-overexpressed IEC models, and the schematic
representation of IEC monolayer grouping. (A) ST3Gal1 mRNA levels
in IEC, sh-Ctr/IEC and ST3Gal1-interfered IEC groups 1–3. (B)
ST3Gal1 mRNA levels in IEC, OE-Ctr/IEC and ST3Gal1-OE/IEC groups.
(C) ST3Gal1 protein levels in IEC, sh-Ctr/IEC and
ST3Gal1-interfered IEC groups. (D) ST3Gal1 protein levels in IEC,
OE-Ctr/IEC and ST3Gal1-OE/IEC groups. (E) Establishment of IEC
monolayers containing sh-Ctr, ST3Gal1-sh3, OE-Ctr and ST3Gal1-OE.
The stable triple culture models were treated with fresh complete
medium supplemented with 2% DSS for 4 days to obtain inflamed
models. Data are presented as the mean ± SD. Statistical
significance was assessed using one-way analysis of variance
followed by Tukey's post-hoc test. **P<0.01. IEC, intestinal
epithelial cell; sh, short hairpin; Ctr, control; OE,
overexpression; PMA, phorbol 12-myristate 13-acetate; DSS, dextran
sulfate sodium.

Figure 3

ST3Gal1 expression in triple culture
models modulates the epithelial barrier of the IEC monolayers. (A)
TEER levels in the IEC monolayers containing sh-Ctr/IEC or
ST3Gal1-sh3/IEC with 2% DSS stimulation (inflamed triple culture)
or without 2% DSS treatment (non-inflamed triple culture). (B) TEER
levels in the inflamed (DSS-treated) and non-inflamed IEC
monolayers containing OE-Ctr/IEC or ST3Gal1-OE/IEC. (C) Results of
the FITC-dextran permeability assay indicated that there was a
significant reduction in the inflamed IEC monolayer containing
ST3Gal1-sh3/IEC. (D) There was a significant increase in the
permeability of the inflamed IEC monolayer containing
ST3Gal1-OE/IEC. Data are presented as the mean ± SD. Statistical
significance was assessed using one-way analysis of variance
followed by Tukey's post-hoc test. **P<0.01. TEER,
trans-epithelial electrical resistance; IEC, intestinal epithelial
cell; sh, short hairpin; Ctr, control; OE, overexpression; DSS,
dextran sulfate sodium.

Figure 4

ST3Gal1 expression in inflamed IEC
monolayers alters MUC2, TFF3, CDX2, STAT3 and p-STAT3 expression.
ST3Gal1 knockdown in ST3Gal1-sh3/IEC significantly increased (A)
MUC2, (B) TFF3 and (C) CDX2, but significantly reduced (D) STAT3
mRNA levels in the inflamed IEC monolayer. ST3Gal1 OE in
ST3Gal1-OE/IEC significantly reduced (E) MUC2, (F) TFF3 and (G)
CDX2 mRNA levels in the inflamed IEC monolayer. (H) ST3Gal1 OE in
ST3Gal1-OE/IEC slightly decreased STAT3 mRNA levels in the inflamed
IEC monolayer. (I) Western blotting showed no changes in protein
expression in non-inflamed knockdown samples. (J) Western blotting
showed that MUC2, TFF3 and CDX2 protein levels were increased, and
p-STAT3 expression was decreased in the inflamed IEC monolayer
comprising ST3Gal1-sh3/IEC. (K) Western blotting showed no notable
changes in protein expression in non-inflamed OE samples. (L)
Western blotting showed that MUC2, TFF3 and CDX2 protein levels
were decreased, and p-STAT3 expression was increased in the
inflamed IEC monolayer comprising ST3Gal1-OE/IEC. Data are
presented as the mean ± SD. Statistical significance was assessed
using one-way analysis of variance followed by Tukey's post-hoc
test. *P<0.05 and **P<0.01. IEC, intestinal epithelial cell;
sh, short hairpin; Ctr, control; OE, overexpression; MUC2, mucin 2;
TFF3, trefoil factor 3; CDX2, homeobox protein CDX-2; p-,
phosphorylated.

Figure 5

Comparison of (A) MUC2, (B) TFF3 and
(C) CDX2 mRNA levels between non-lesional mucosa (n=44) and
lesional mucosa (n=75) from distinct patients with ulcerative
colitis. Correlations between (D) MUC2, (E) TFF3 and (F) CDX2 mRNA
levels and ST3Gal1 expression. Data in A-C are presented as box and
whisker plots, and those in D-F as scatter plots. Statistical
significance for group comparisons in A-C was assessed using
unpaired t-test. Correlation analyses in D-F were performed using
Pearson's correlation coefficient. *P<0.05 and **P<0.01. Data
were obtained from the GSE107499 dataset from the Gene Expression
Omnibus database. *P<0.05 and **P<0.01. NS, not significant;
MUC2, mucin 2; TFF3, trefoil factor 3; CDX2, homeobox protein
CDX-2.

Figure 6

ST3Gal1 expression in the IEC
monolayer alters the levels of inflammatory mediators. ST3Gal1
knockdown in ST3Gal1-sh3/IEC significantly reduced the mRNA levels
of the inflammatory mediators (A) IL-1β, (B) IL-6 and (C) IL-8 from
the inflamed IEC monolayer supernatant. ST3Gal1 OE in
ST3Gal1-OE/IEC significantly increased the mRNA levels of the
inflammatory mediators (D) IL-1β, (E) IL-6 and (F) IL-8 in the
supernatant of the inflamed IEC monolayer. Protein levels of the
inflammatory mediators (G) IL-1β, (H) IL-6 and (I) IL-8 were
decreased in the supernatant of inflamed ST3Gal1-sh3/IEC. Protein
levels of the inflammatory mediators (J) IL-1β, (K) IL-6 and (L)
IL-8 were increased in the supernatant of inflamed ST3Gal1-OE/IEC.
Data are presented as the mean ± SD, and statistical significance
was assessed using one-way ANOVA followed by Tukey's post-hoc test.
**P<0.01. IEC, intestinal epithelial cell; sh, short hairpin;
Ctr, control; OE, overexpression.

Figure 7

Comparison of the expression of
inflammatory mediators (A) IL-1β, (B) IL-6 and (C) IL-8, as well as
(D) STAT3 between non-lesional mucosa (n=44) and lesional mucosa
(n=75) from distinct patients with ulcerative colitis. Correlations
between (E) IL-1β, (F) IL-6, (G) IL-8 and (H) STAT3 mRNA levels and
ST3Gal1 expression. Data were obtained from the GSE107499 dataset
from the Gene Expression Omnibus database. Data in A-D are
presented as box and whisker plots, and those in E-H as scatter
plots. Statistical significance for group comparisons in A-D was
assessed using the unpaired t-test; correlation analyses in E-H
were performed via Pearson's correlation coefficient.
**P<0.01.
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Copy and paste a formatted citation
Spandidos Publications style
Tian Y, Liu Y, Shang Y, Ran L, Liu L, Wang R and Ye J: ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis. Mol Med Rep 33: 73, 2026.
APA
Tian, Y., Liu, Y., Shang, Y., Ran, L., Liu, L., Wang, R., & Ye, J. (2026). ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis. Molecular Medicine Reports, 33, 73. https://doi.org/10.3892/mmr.2025.13783
MLA
Tian, Y., Liu, Y., Shang, Y., Ran, L., Liu, L., Wang, R., Ye, J."ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis". Molecular Medicine Reports 33.2 (2026): 73.
Chicago
Tian, Y., Liu, Y., Shang, Y., Ran, L., Liu, L., Wang, R., Ye, J."ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis". Molecular Medicine Reports 33, no. 2 (2026): 73. https://doi.org/10.3892/mmr.2025.13783
Copy and paste a formatted citation
x
Spandidos Publications style
Tian Y, Liu Y, Shang Y, Ran L, Liu L, Wang R and Ye J: ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis. Mol Med Rep 33: 73, 2026.
APA
Tian, Y., Liu, Y., Shang, Y., Ran, L., Liu, L., Wang, R., & Ye, J. (2026). ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis. Molecular Medicine Reports, 33, 73. https://doi.org/10.3892/mmr.2025.13783
MLA
Tian, Y., Liu, Y., Shang, Y., Ran, L., Liu, L., Wang, R., Ye, J."ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis". Molecular Medicine Reports 33.2 (2026): 73.
Chicago
Tian, Y., Liu, Y., Shang, Y., Ran, L., Liu, L., Wang, R., Ye, J."ST3Gal1 modulates intestinal barrier function and impacts human ulcerative colitis". Molecular Medicine Reports 33, no. 2 (2026): 73. https://doi.org/10.3892/mmr.2025.13783
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