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MEIS1‑regulated miR‑488‑3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C

  • Authors:
    • Chunming Zhang
    • Wenjing Hao
    • Xinfang Wang
    • Huina Guo
    • Long He
    • Jiao Yang
    • Ying Wang
    • Xiwang Zheng
    • Zhongxun Li
    • Qi Han
    • Liqi Wen
    • Hongliang Liu
  • View Affiliations / Copyright

    Affiliations: Shanxi Key Laboratory of Otorhinolaryngology Head and Neck Cancer, First Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, Department of Anatomy, The Basic Medical School of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China
    Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 142
    |
    Published online on: July 11, 2025
       https://doi.org/10.3892/ijmm.2025.5583
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Abstract

Laryngeal squamous cell carcinoma (LSCC) is a common malignant tumor originating from the mucosal epithelium of the larynx. MicroRNA (miR)‑488‑3p has non‑negligible multifaceted roles in some types of cancer; however, its association with LSCC has not yet been reported. Our prior RNA sequencing data indicated that miR‑488‑3p expression is downregulated in LSCC tissue, yet the detailed function and regulatory mechanism of miR‑488‑3p in LSCC remain unknown. In the present study, quantitative PCR analysis corroborated the significant downregulation of miR‑488‑3p in LSCC tumor tissues, with this downregulation being strongly associated with malignant progression in LSCC. Furthermore, overexpression of miR‑488‑3p suppressed LSCC cell proliferation, colony formation, migration, invasion, xenograft tumor growth and epithelial‑mesenchymal transition. Mechanistically, miR‑488‑3p directly interacted with the 3' untranslated region of activin A receptor type 1C (ACVR1C) and downregulated ACVR1C expression. Functional experiments revealed that miR‑488‑3p suppressed the malignant phenotypes of LSCC via ACVR1C. Additionally, bioinformatics analysis coupled with chromatin immunoprecipitation assay revealed that myeloid ecotropic viral integration site 1 (MEIS1) promoted the expression of miR‑488‑3p transcriptionally by directly binding its promoter region. Collectively, the results demonstrated that miR‑488‑3p acts as a tumor suppressor molecule in LSCC, and a role was established for the MEIS1/miR‑488‑3p/ACVR1C axis in regulating LSCC progression, thus providing novel potential biomarkers and targets for patients with LSCC.
View Figures

Figure 1

Downregulation of miR-488-3p and its
clinical significance in LSCC. The expression levels of miR-488-3p
were detected by qPCR in 65 paired LSCC and ANM samples. (A)
Comparison of miR-488-3p expression between LSCC and matched ANM
samples. (B) Expression levels of miR-488-3p in 65 paired LSCC and
matched ANM tissues. Analysis of association between miR-488-3p
expression and (C) T stages, (D) clinical stages, (E) lymph node
metastasis (N stages) and (F) degree of differentiation. (G) qPCR
analysis of miR-488-3p expression in human LSCC and normal cell
lines. A comparison of miR-488-3p expression in LSCC FD-LSC-1 and
AMC-HN-8 cells with normal revealed significantly lower miR-488-3p
expression in LSCC cells. ***P<0.001 vs. 2BS cells.
miR, microRNA; LSCC, laryngeal squamous cell carcinoma; qPCR,
quantitative PCR; ANM, adjacent normal mucosa; N.S., not
significant.

Figure 2

miR-488-3p overexpression inhibits
LSCC cell proliferation, migration, invasion and
epithelial-mesenchymal transition. (A) Quantitative PCR analysis of
miR-488-3p expression in LSCC cells transfected with miR-488-3p or
NC mimics. (B) Cell Counting Kit-8 and (C) colony formation assays
detected the proliferative ability of LSCC cells transfected with
miR-488-3p or NC mimics. Transwell assays determined the (D)
migration and (E) invasion of LSCC cells transfected with
miR-488-3p or NC mimics. (F) Western blotting of the protein levels
of E-cadherin, N-cadherin and vimentin. Cropped images represent
different blots, but samples are from the same experiment and the
blots were processed in parallel. Scale bars, 100 µm.
*P<0.05, **P<0.01 and
***P<0.001. miR, microRNA; LSCC, laryngeal squamous
cell carcinoma; NC, negative control.

Figure 3

ACVR1C is a direct target gene of
miR-488-3p. (A) Venn analysis of the target genes of miR-488-3p
predicted by TargetScan, miRDB and miRWalk. (B) Venn analysis of
the potential target genes of miR-488-3p and upregulated mRNAs in
LSCC. (C) Quantitative PCR analysis of the mRNA levels of ACVR1C,
ABCA12, TNFSF11 and SYT14 in LSCC cells transfected with miR-488-3p
or NC mimics. (D) Western blotting of the protein levels of ACVR1C.
(E) Schematic of ACVR1C 3′ UTR sequence containing WT and Mut
miR-488-3p binding sites. Luciferase reporter assay measured the
luciferase activity of ACVR1C 3′ UTR when FD-LSC-1 cells were
transfected with miR-488-3p (F) mimics or (G) inhibitor.
**P<0.01 and ***P<0.001. ACVR1C,
activin A receptor type 1C; miR, microRNA; LSCC, laryngeal squamous
cell carcinoma; NC, negative control; UTR, untranslated region; WT,
wild type; Mut, mutant; N.S., not significant.

Figure 4

ACVR1C knockdown inhibits LSCC cell
proliferation, migration, invasion and epithelial-mesenchymal
transition. (A) Analysis of ACVR1C expression in RNA-seq data
(GSE127165). (B) Quantitative PCR and (C) western blot analysis of
the mRNA and protein expression levels of ACVR1C in AMC-HN-8 and
FD-LSC-1 cells transfected with si-ACVR1C-1 and 2 or si-NC. Cropped
images represent different blots, but samples are from the same
experiment and the blots were processed in parallel. (D) Cell
Counting Kit-8 and (E) colony formation assays investigated the
proliferative ability of LSCC cells transfected with si-ACVR1C or
si-NC. In the Transwell assay, ACVR1C knockdown significantly
suppressed the (F) migration and (G) invasion of LSCC cells. (H)
Western blot analysis of the protein levels of E-cadherin,
N-cadherin and vimentin in LSCC cells transfected with si-ACVR1C or
si-NC. Cropped images represent different blots, but samples are
from the same experiment and the blots were processed in parallel.
Scale bar, 100 µm. *P<0.05,
**P<0.01 and ***P<0.001. ACVR1C,
activin A receptor type 1C; LSCC, laryngeal squamous cell
carcinoma; NC, negative control; si, small interfering RNA.

Figure 5

miR-488-3p suppresses LSCC cell
proliferation, migration and invasion by targeting ACVR1C. (A)
Western blot analysis of ACVR1C protein in FD-LSC-1 and AMC-HN-8
cells transfected with OE-ACVR1C or control empty plasmid (OE-NC).
Flag antibodies were used to detect the Flag-tagged ACVR1C
proteins. (B) Western blot analysis of ACVR1C protein expression in
FD-LSC-1 and AMC-HN-8 cells co-transfected with OE-NC and NC mimics
(Control), OE-NC and miR-488-3p mimics (miR-488-3p mimics), NC
mimics and ACVR1C OE (ACVR1C OE) or miR-488-3p mimics and ACVR1C OE
(miR-488-3p mimics + ACVR1C OE). (C) Cell Counting Kit-8 and (D)
colony formation assays detected the proliferative ability of LSCC
cells in different groups. (E and F) Transwell assays determined
the migration and invasion of LSCC cells in different groups. Scale
bar, 100 µm. **P<0.01 and
***P<0.001. ACVR1C, activin A receptor type 1C; miR,
microRNA; LSCC, laryngeal squamous cell carcinoma; OE,
overexpression.

Figure 6

MEIS1 directly binds to the promoter
region of miR-488 and upregulates the expression level of
miR-488-3p. (A) Schematic of the core region of the miR-488
promoter and its potential MEIS1-binding sites. The binding sites
for MEIS1 in the miR-488 promoter region were identified using the
LASAGNA-Search 2.0 web tool and the JASPAR2024 database.
The MEIS1-binding motif was also taken from the
JASPAR2024 database. Primers were used to detect levels
of miR-488 promoter fragments directly bound to MEIS1 in the ChIP
assay. (B) ChIP-qPCR assay evaluated MEIS1-binding site
interactions in the miR-488 promoter region. (C) Targeted
relationship of MEIS1 to the miR-488 promoter was validated in 293T
cells using dual-luciferase reporter gene assay. (D) Analysis of
MEIS1 expression in RNA-seq data (GSE127165). (E) Correlation
analysis of MEIS1 and miR-488-3p expression using RNA-seq data
(GSE127165, GSE133632). (F) qPCR analysis of miR-488-3p expression
in LSCC cells transfected with OE-MEIS1 or OE-NC.
**P<0.01 and ***P<0.001. MEIS1, myeloid
ecotropic viral integration site 1; N.S., not significant; miR,
microRNA; qPCR, quantitative PCR; OE, overexpression; NC, negative
control; LSCC, laryngeal squamous cell carcinoma; ChIP, chromatin
immunoprecipitation.

Figure 7

Downregulation of miR-488-3p
partially reverses MEIS1-mediated tumor suppression. (A) Western
blot analysis of MEIS1 protein in FD-LSC-1 and AMC-HN-8 cells
transfected with OE-MEIS1 or co-transfected with OE-MEIS1 and
miR-488-3p inhibitor. Flag antibodies were used to detect the
Flag-tagged MEIS1 proteins. (B) qPCR analysis of miR-488-3p
expression in LSCC cells transfected with miR-488-3p or NC
inhibitors. (C) Cell Counting Kit-8 and (D) colony formation assays
estimated the proliferative ability of LSCC cells in different
groups. (E and F) Transwell assays determined the migration and
invasion of LSCC cells in different groups. Scale bar, 100
µm. *P<0.05, **P<0.01 and
***P<0.001. MEIS1, myeloid ecotropic viral
integration site 1; miR, microRNA; OE, overexpression; NC, negative
control; LSCC, laryngeal squamous cell carcinoma.

Figure 8

Effect of miR-488-3p on tumor growth
in vivo. (A) Image of xenograft tumors after intratumoral
injection of miR-488-3p or NC agomir. The xenograft tumor volume
was measured to plot the growth curve. (B) Final weight of
xenograft tumors. (C) Quantitative PCR analysis of miR-488-3p
expression in xenograft tumors. (D) Representative
immunohistochemical staining of vimentin, N-cadherin, E-cadherin
and Ki-67 expression in xenograft tumors. (E) Schematic
representation of the MEIS1/miR-488-3p/ACVR1C axis in regulating
laryngeal squamous cell carcinoma progression. The red and green
arrows indicate upregulation and downregulation in LSCC,
respectively. Scale bar, 50 mm. *P<0.05,
**P<0.01 and ***P<0.001. miR, microRNA;
NC, negative control; MEIS1, myeloid ecotropic viral integration
site 1; ACVR1C, activin A receptor type 1C.
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Copy and paste a formatted citation
Spandidos Publications style
Zhang C, Hao W, Wang X, Guo H, He L, Yang J, Wang Y, Zheng X, Li Z, Han Q, Han Q, et al: MEIS1‑regulated miR‑488‑3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C. Int J Mol Med 56: 142, 2025.
APA
Zhang, C., Hao, W., Wang, X., Guo, H., He, L., Yang, J. ... Liu, H. (2025). MEIS1‑regulated miR‑488‑3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C. International Journal of Molecular Medicine, 56, 142. https://doi.org/10.3892/ijmm.2025.5583
MLA
Zhang, C., Hao, W., Wang, X., Guo, H., He, L., Yang, J., Wang, Y., Zheng, X., Li, Z., Han, Q., Wen, L., Liu, H."MEIS1‑regulated miR‑488‑3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C". International Journal of Molecular Medicine 56.3 (2025): 142.
Chicago
Zhang, C., Hao, W., Wang, X., Guo, H., He, L., Yang, J., Wang, Y., Zheng, X., Li, Z., Han, Q., Wen, L., Liu, H."MEIS1‑regulated miR‑488‑3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C". International Journal of Molecular Medicine 56, no. 3 (2025): 142. https://doi.org/10.3892/ijmm.2025.5583
Copy and paste a formatted citation
x
Spandidos Publications style
Zhang C, Hao W, Wang X, Guo H, He L, Yang J, Wang Y, Zheng X, Li Z, Han Q, Han Q, et al: MEIS1‑regulated miR‑488‑3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C. Int J Mol Med 56: 142, 2025.
APA
Zhang, C., Hao, W., Wang, X., Guo, H., He, L., Yang, J. ... Liu, H. (2025). MEIS1‑regulated miR‑488‑3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C. International Journal of Molecular Medicine, 56, 142. https://doi.org/10.3892/ijmm.2025.5583
MLA
Zhang, C., Hao, W., Wang, X., Guo, H., He, L., Yang, J., Wang, Y., Zheng, X., Li, Z., Han, Q., Wen, L., Liu, H."MEIS1‑regulated miR‑488‑3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C". International Journal of Molecular Medicine 56.3 (2025): 142.
Chicago
Zhang, C., Hao, W., Wang, X., Guo, H., He, L., Yang, J., Wang, Y., Zheng, X., Li, Z., Han, Q., Wen, L., Liu, H."MEIS1‑regulated miR‑488‑3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C". International Journal of Molecular Medicine 56, no. 3 (2025): 142. https://doi.org/10.3892/ijmm.2025.5583
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