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Article Open Access

lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating β‑catenin via enhancing H3K18 lactylation

  • Authors:
    • Li Yu
    • Nian Zhou
    • Xiao-Liang Zhang
    • Xue-Jing Pang
    • Lu Xing
    • Yun-Jing Pu
    • Li Zhang
    • Jing-Nan Wu
    • Hong Shu
  • View Affiliations / Copyright

    Affiliations: Department of Dermatology, Kunming Children's Hospital (Children's Hospital Affiliated to Kunming Medical University), Kunming, Yunnan 650228, P.R. China, Department of Orthopedics, Kunming Municipal Hospital of Traditional Chinese Medicine, Kunming, Yunnan 650011, P.R. China, Department of Stomatology, Kunming Children's Hospital (Children's Hospital Affiliated to Kunming Medical University), Kunming, Yunnan 650228, P.R. China
    Copyright: © Yu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 53
    |
    Published online on: November 25, 2025
       https://doi.org/10.3892/mmr.2025.13763
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Abstract

Infantile hemangioma (IH), a common vascular tumor, occurs in childhood; however, its pathogenesis has not been fully elucidated. In the present study, the roles and detailed mechanisms of long non‑coding RNA (lncRNA) NEAT1 in the progression of hemangioma were further explored. The NEAT1‑interacting proteins were selected by analyzing the catRAPID database and lactate dehydrogenase B (LDHB) was predicted to bind with NEAT1. The binding between NEAT1 and LDHB was validated using an RNA immunoprecipitation assay and it was further found that knocking down NEAT1 expression destabilized LDHB by regulating the proteasome pathway. The knocking down of lncRNA NEAT1 also inhibited cellular protein lactylation and downregulated β‑catenin. Furthermore, blockade of lactylation via 2‑DG and oxamate attenuated the viability and colony formation of hemangioma cells. NEAT1 promoted the lactylation of H3K18 in the promoter region of β‑catenin, and blockade of lactylation downregulated β‑catenin expression in hemangioma cells. The lactyltransferases alanyl‑tRNA synthetase 1 and P300 were regulated by NEAT1 and also positively regulated β‑catenin. The levels of β‑catenin mRNA and H3K18 lactylation were also found to be elevated in IH tissues. Taken together, the results of the present study revealed that lncRNA NEAT1, which is upregulated in hemangioma, binds with and stabilizes LDHB, subsequently elevates the levels of cellular lactate and H3K18 lactylation, potentiates β‑catenin transcription and ultimately enhances the proliferation of hemangioma cells.
View Figures

Figure 1

Identification of proteins that
interact with the long non-coding RNA NEAT1. (A) BP enrichment
analysis of NEAT1-interacting proteins. (B) CC enrichment analysis
of NEAT1-interacting proteins. (C) MF enrichment analysis of
NEAT1-interacting proteins. (D) KEGG enrichment analysis of
NEAT1-interacting proteins. BP, Biological Process; CC, Cellular
Component; MF, Molecular Function; KEGG, Kyoto Encyclopedia of
Genes and Genomes.

Figure 2

Long non-coding RNA NEAT1 interacts
with and positively regulates LDHB in hemangioma cells. (A) The
interaction propensity between NEAT1 and LDHB was predicted by the
catRAPID software. (B) The interaction map between NEAT1 and LDHB
was predicted by the catRAPID software. (C) The binding between
NEAT1 and LDHB was measured by the RNA immunoprecipitation assay;
****P<0.0001 (n=3). (D) The knockdown efficiency of NEAT1 was
detected by RT-qPCR assay; **P<0.01 vs. NC (n=3). (E) The
protein level of LDHB in NEAT1-knock down cells was detected by
western blotting; ****P<0.0001 vs. NC (n=3). (F) The LDHB mRNA
levels in NEAT1-knock down cells were detected by RT-qPCR (n=3).
(G) After CHX treatment, the protein expression level of LDHB in
NEAT1-knock down cells was detected by western blotting;
**P<0.01, ***P<0.001 (n=3). (H) The proteasome inhibitor
MG132 and the lysosome inhibitor BafA1 were used to treat the
NEAT1-knock down cells, and the protein expression level of LDHB
was detected by western blotting. ns, not significant; CHX,
cycloheximide; BafA1, bafilomycin A1; NC, negative control; si,
small interfering (RNA); LDHB, lactate dehydrogenase B.

Figure 3

Knocking down long non-coding RNA
NEAT1 inhibits cellular protein lactylation and downregulates
β-catenin in hemangioma cells. (A) Cellular lactate levels were
measured after NEAT1 knockdown; ****P<0.0001 vs. NC (n=3). (B)
The level of pan-lactylation in NEAT1-knock down cells were
detected by western blotting (n=3). (C) H3K18 lactylation in
NEAT1-knock down cells was detected by western blotting (n=3). (D)
Quantitative analysis of the levels of H3K18la in the western
blotting results; *P<0.05, **P<0.01 vs. NC (n=3). (E) The
knockdown efficiency of LDHB was detected by western blotting;
*P<0.05 (n=3). (F) The viability of LDHB-knock down cells was
detected via Cell Counting Kit-8 assay; **P<0.01 (n=3). (G)
Colony formation ability of LDHB-knock down cells was detected via
a colony formation assay; **P<0.01 (n=3). (H) The mRNA levels of
CTNNB1 in NEAT1-knock down cells were detected using
RT-qPCR. ****P<0.0001 vs. NC (n=3). (I) The mRNA levels of
CTNNB1 in LDHB-knock down cells were detected using RT-qPCR.
***P<0.001 vs. NC (n=3). (J) The protein levels of β-catenin in
NEAT1-knock down and LDHB-knock down cells were measured using
western blotting (n=3). (K) Semi-quantitative analysis of the
levels of β-catenin in NEAT1-knock down cells detected by western
blotting; ***P<0.001 vs. NC (n=3). (L) Semi-quantitative
analysis of the levels of β-catenin in LDHB-knock down cells
detected by western blotting; ***P<0.001 vs. NC (n=3). NC,
negative control; H3K18la, Histone H3K18 lactylation; si, small
interfering (RNA); LDHB, lactate dehydrogenase B; CTNNB1, β-catenin
gene; RT-qPCR, reverse transcription-quantitative polymerase chain
reaction.

Figure 4

Blockade of lactylation using 2-DG
and oxamate attenuates the viability and colony formation of
hemangioma cells, and lactate treatment has the opposite effects.
(A) 2-DG and oxamate suppress lactate production by blocking
glycolysis. (B) Western blotting was used to detect pan-lac in
hemangioma cells after 2-DG and oxamate treatment. (C) Western
blotting was used to detect the pan-lactylation in hemangioma cells
after the addition of lactate. The viability of hemangioma cells
under (D) 2-DG, (E) oxamate and (F) lactate treatment;
***P<0.001 (n=3). Colony formation ability of hemangioma cells
under (G) 2-DG, (H) oxamate and (I) lactate treatment; **P<0.01,
***P<0.001 (n=3). 2-DG, 2-Deoxy-D-glucose; LDH, lactate
dehydrogenase; pan-lac, pan-lactylation; H3K18la, Histone H3K18
lactylation.

Figure 5

Long non-coding RNA NEAT1 promotes
the H3K18 lactylation of the CTNNB1 promoter and the
blockade of lactylation downregulates β-catenin in hemangioma
cells. H3K18 lactylation levels of the CTNNB1 promoter in
(A) 2-DG- or oxamate-treated cells and (B) lactate-treated, (C)
NEAT1-knock down and (D) LDHB-knock down hemangioma cells were
evaluated using ChIP-PCR; *P<0.05, **P<0.01, ***P<0.001
vs. NC (n=3). The CTNNB1 mRNA levels in (E) 2-DG- and
oxamate-treated cells as well as (F) lactate-treated cells were
detected by RT-qPCR; **P<0.01, ****P<0.0001 vs. NC (n=3). The
protein levels of β-catenin in (G) 2-DG- and oxamate-treated cells
were detected by western blotting (n=3). (H) Semi-quantitative
analysis of the levels of β-catenin in 2-DG and oxamate-treated
cells detected by western blotting; ***P<0.001 vs. NC (n=3). (I)
The protein levels of β-catenin in lactate-treated cells were
detected by western blotting (n=3). (J) Semi-quantitative analysis
of the levels of β-catenin in lactate-treated cells detected by
western blotting; *P<0.05 vs. NC (n=3). 2-DG, 2-Deoxy-D-glucose;
NC, negative control; CTNNB1, β-catenin gene; LDHB, lactate
dehydrogenase B; ChIP, chromatin immunoprecipitation; RT-qPCR,
reverse transcription-quantitative polymerase chain reaction; si,
small interfering (RNA).

Figure 6

Long non-coding RNA NEAT1 regulates
β-catenin by positively affecting the lactyltransferases AARS1 and
P300 in hemangioma cells. (A) The knockdown efficiencies of AARS1,
AARS2, CBP, KAT5, KAT8 and P300 were confirmed by RT-qPCR;
****P<0.0001 vs. NC (n=3). (B) After AARS1, AARS2, CBP, KAT5,
KAT8 and P300 were knocked down, the CTNNB1 mRNA level was
detected using RT-qPCR; *P<0.05, ***P<0.001, ****P<0.0001
vs. NC (n=3). (C) The knockdown efficiencies of SIRT1, SIRT2,
SIRT3, HDAC1, HDAC2 and HDAC3 were confirmed by RT-qPCR;
****P<0.0001 vs. NC (n=3). (D) After SIRT1, SIRT2, SIRT3, HDAC1,
HDAC2 and HDAC3 were knocked down, the CTNNB1 mRNA level was
detected using RT-qPCR; ****P<0.0001 vs. NC (n=3). (E) The
knockdown efficiency of AARS1 and P300 was confirmed by western
blotting (n=3). (F) Semi-quantitative analysis of the levels of
AARS1 and P300 in AARS1-knock down and P300-knock down cells
detected by western blotting; **P<0.01, ****P<0.0001 vs. NC
(n=3). (G) The protein level of β-catenin after AARS1 and P300
knockdown was detected using western blotting (n=3). (H)
Semi-quantitative analysis of the levels of β-catenin in
AARS1-knock down and P300-knock down cells detected by western
blotting; **P<0.01, ***P<0.001 vs. NC (n=3). (I) The mRNA
levels of AARS1 and P300 after NEAT1 knockdown were detected using
RT-qPCR; ****P<0.0001 vs. NC (n=3). (J) The protein levels of
AARS1 and P300 after NEAT1 knockdown were detected using western
blotting (n=3). (K) Semi-quantitative analysis of the levels of
AARS1 and P300 in NEAT1-knock down cells detected by western
blotting; **P<0.01, ***P<0.001, ****P<0.0001 vs. NC (n=3).
ns, not significant; NC, negative control; AARS, alany-tRNA
synthetase; CBP, CREB binding lysine acetyltransferase; KAT, lysine
acetyltransferase; RT-qPCR, reverse transcription-quantitative
polymerase chain reaction; CTNNB1, β-catenin gene; SIRT, sirtuin;
HDAC, histone deacetylase; siRNA, small interfering RNA.

Figure 7

β-catenin and H3K18 lactylation
levels were elevated in IH tissues. (A) The viability of hemangioma
cells treated with a P300 inhibitor (A-485) were measured via Cell
Counting Kit-8 assay; ***P<0.001 (n=3). (B) The colony formation
ability of hemangioma cells treated with a P300 inhibitor (A-485)
were measured via colony formation assays (n=3). (C) Quantitative
analysis of the colony formation results; **P<0.01 (n=3). (D)
The CTNNB1 mRNA level after A-485 treatment was detected
using RT-qPCR; **P<0.01 (n=3). (E) The protein level of
β-catenin after A-485 treatment was detected using western blotting
(n=3). (F) Semi-quantitative analysis of the levels of β-catenin in
A-485-treated cells detected by western blotting; **P<0.01
(n=3). (G) The CTNNB1 mRNA level in IH tissues was detected
using RT-qPCR; **P<0.01 (n=10). (H) The level of β-catenin
protein in IH tissues was detected using western blotting. (I) The
level of H3K18 lactylation in IH tissues was detected using western
blotting. (J) Proposed model of long non-coding RNA NEAT1
regulation of the LDHB/H3K18-lactylation/β-catenin signaling
cascade. IH, infantile hemangioma; NC, negative control; CTNNB1,
β-catenin gene; RT-qPCR, reverse transcription-quantitative
polymerase chain reaction; AARS1, alany-tRNA synthetase 1; LDHB,
lactate dehydrogenase B.
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Copy and paste a formatted citation
Spandidos Publications style
Yu L, Zhou N, Zhang X, Pang X, Xing L, Pu Y, Zhang L, Wu J and Shu H: lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating &beta;‑catenin via enhancing H3K18 lactylation. Mol Med Rep 33: 53, 2026.
APA
Yu, L., Zhou, N., Zhang, X., Pang, X., Xing, L., Pu, Y. ... Shu, H. (2026). lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating &beta;‑catenin via enhancing H3K18 lactylation. Molecular Medicine Reports, 33, 53. https://doi.org/10.3892/mmr.2025.13763
MLA
Yu, L., Zhou, N., Zhang, X., Pang, X., Xing, L., Pu, Y., Zhang, L., Wu, J., Shu, H."lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating &beta;‑catenin via enhancing H3K18 lactylation". Molecular Medicine Reports 33.2 (2026): 53.
Chicago
Yu, L., Zhou, N., Zhang, X., Pang, X., Xing, L., Pu, Y., Zhang, L., Wu, J., Shu, H."lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating &beta;‑catenin via enhancing H3K18 lactylation". Molecular Medicine Reports 33, no. 2 (2026): 53. https://doi.org/10.3892/mmr.2025.13763
Copy and paste a formatted citation
x
Spandidos Publications style
Yu L, Zhou N, Zhang X, Pang X, Xing L, Pu Y, Zhang L, Wu J and Shu H: lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating &beta;‑catenin via enhancing H3K18 lactylation. Mol Med Rep 33: 53, 2026.
APA
Yu, L., Zhou, N., Zhang, X., Pang, X., Xing, L., Pu, Y. ... Shu, H. (2026). lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating &beta;‑catenin via enhancing H3K18 lactylation. Molecular Medicine Reports, 33, 53. https://doi.org/10.3892/mmr.2025.13763
MLA
Yu, L., Zhou, N., Zhang, X., Pang, X., Xing, L., Pu, Y., Zhang, L., Wu, J., Shu, H."lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating &beta;‑catenin via enhancing H3K18 lactylation". Molecular Medicine Reports 33.2 (2026): 53.
Chicago
Yu, L., Zhou, N., Zhang, X., Pang, X., Xing, L., Pu, Y., Zhang, L., Wu, J., Shu, H."lncRNA NEAT1 promotes the proliferation of hemangioma cells by transcriptionally activating &beta;‑catenin via enhancing H3K18 lactylation". Molecular Medicine Reports 33, no. 2 (2026): 53. https://doi.org/10.3892/mmr.2025.13763
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