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Article Open Access

METTL5‑mediated m6A modification of 18S rRNA drives oral squamous cell carcinoma progression by enhancing CCND3 translation

  • Authors:
    • Lianlian Liu
    • Surui Sheng
    • Xin Zheng
    • Shuojin Huang
    • Lu Wang
    • Dalong Shu
    • Leile Tang
    • Qianting He
  • View Affiliations / Copyright

    Affiliations: Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Sun Yat‑Sen University, Guangzhou, Guangdong 510080, P.R. China, Department of Oral and Maxillofacial‑Head and Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China, Department of Medical Laboratory, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Cardiovascular Medicine, The Third Affiliated Hospital, Sun Yat‑Sen University, Guangzhou, Guangdong 510630, P.R. China
    Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 137
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    Published online on: February 12, 2026
       https://doi.org/10.3892/ol.2026.15490
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Abstract

Oral squamous cell carcinoma (OSCC) is a predominant head and neck malignancy with epigenetic alterations key to its development; however, the role and mechanisms of methyltransferase 5, N6‑adenosine (METTL5) in OSCC remain to be elucidated. This study aimed to investigate the functional role and underlying mechanism of METTL5 in OSCC progression. Western blotting analyzed METTL5 expression in OSCC cell lines and tissues, whilst Cell Counting Kit‑8, Transwell, colony formation and wound healing assays, as well as cell cycle analysis and nude mouse xenografts were used to assess its functional effects. Moreover, ribosome nascent‑chain complex‑bound messenger RNA sequencing (RNC‑seq) was used to assess gene translation efficiency (TE). Western blotting, Transwell and colony formation assays, and cell cycle analysis were employed for downstream studies. Bioinformatics analyses were conducted to complement mechanistic exploration. The Cancer Genome Atlas data demonstrated that higher METTL5 expression in metastatic head and neck squamous cell carcinoma tissues was significantly associated with poor prognosis. Furthermore, METTL5 knockout significantly inhibited OSCC tumorigenesis and metastasis in vitro and in vivo. Finally, RNC‑seq identified 3,391 genes with reduced TE, with cyclin D3 (CCND3) shown to be a downstream target. In conclusion, METTL5 may promote OSCC progression by regulating CCND3 via N6‑methyladenosine‑mediated ribosomal methylation, positioning it as a potential therapeutic target in the future.
View Figures

Figure 1

METTL5 is elevated in HNSCC and is
associated with poor prognosis of HNSCC. (A) Comparison of mRNA
levels of METTL5 in HNSCC with normal tissues in TCGA-HNSCC
dataset. (B) Overall survival of patients between METTL5-high
(n=259) and METTL5-low (n=259) groups. (C) METTL5 expression level
in normal and HNSCC tissues with different stages of lymph node
metastasis. (D) METTL5 expression level in normal tissues and
different stages of HNSCC tissues. (E) METTL5 expression level in
normal HNSCC tissues and different grades of HNSCC tissues. (F)
METTL5 protein level in OSCC(HSC3, SCC15) and HOK cell lines. (G)
Representative bands of METTL5 protein expression levels in human
OSCC and paired normal tissues measured by western blotting. (H)
Semi-quantification of METTL5 protein expression levels in human
OSCC and paired normal tissues. *P<0.05, **P<0.01;
***P<0.001. METTL5, methyltransferase 5, N6-adenosine; HNSCC,
head and neck squamous cell carcinoma; TCGA, The Cancer Genome
Atlas; OSCC, oral squamous cell carcinoma; HOK, human normal oral
epithelial keratinocyte; HR, hazard ratio; TPM, transcripts per
million; N, normal; T, tumor.

Figure 2

Knockout of METTL5 inhibits the
progression of OSCC in vitro. (A) Validation of METTL5
stable knockout in SCC15 cells by western blotting. Results of (B)
Cell Counting Kit-8 assay, (C) colony formation assay, (D)
Transwell assay and (E) wound healing assay after knockout of
METTL5 in SCC15 cells. *P<0.05, **P<0.01; ***P<0.001;
****P<0.0001. METTL5, methyltransferase 5, N6-adenosine; NC,
negative control; OSCC, oral squamous cell carcinoma; sg, small
guide RNA; NC, negative control; OD, optical density.

Figure 3

Overexpression of METTL5 promotes the
progression of OSCC. (A) Validation of METTL5 overexpression in
SCC15 cells by western blotting. (B) Results of the Cell Counting
Kit-8 assay after METTL5 overexpression in SCC15 cells. (C)
Statistical results of the Transwell assay after METTL5
overexpression in SCC15 cells. (D) Results of the colony formation
assay after knockout of METTL5 in SCC15 cells. **P<0.01. METTL5,
methyltransferase 5, N6-adenosine; OSCC, oral squamous cell
carcinoma; oe, overexpression; NC, negative control; OD, optical
density.

Figure 4

Inhibition of METTL5 suppresses tumor
growth in vivo. (A) Representative images of subcutaneous
tumors from the SCC15 cell mouse model. (B) Quantification of tumor
weight and tumor volume. (C) Tumor volume changes during treatment.
(D) Quantitative analysis of IHC scores for METTL5 and Ki-67
expression. (E) Representative images of METTL5 and Ki-67 staining.
**P<0.01; ***P<0.001. METTL5, methyltransferase 5,
N6-adenosine; sg, small guide RNA; NC, negative control; IHC,
immunohistochemistry.

Figure 5

Knockout of METTL5 selectively
inhibits the translation of oncogenic mRNAs. (A) TE scatter plot of
METTL5 knockout and control SCC15 cells. (B) Pathway enrichment of
TE-downregulated genes. (C) Western blotting confirmed the
decreased protein levels of CCND3 in METTL5 knockout SCC15 cells.
METTL5, methyltransferase 5, N6-adenosine; TE, translation
efficiency; CCND3, cyclin D3; sg, small guide RNA; NC, negative
control.

Figure 6

CCND3 overexpression partially
rescues cancer cell phenotype in METTL5-depleted oral squamous cell
carcinoma cells. (A) Validation of CCND3 overexpression in
METTL5-depleted HSC3 cells. Results of the (B) Transwell, (C)
colony formation and (D) cell cycle assays after CCND3
overexpression in METTL5-depleted HSC3 cells. *P<0.05,
**P<0.01; ***P<0.001; ****P<0.0001. METTL5,
methyltransferase 5, N6-adenosine; CCND3, cyclin D3; sg, small
guide RNA; NC, negative control; ns, not significant.
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Copy and paste a formatted citation
Spandidos Publications style
Liu L, Sheng S, Zheng X, Huang S, Wang L, Shu D, Tang L and He Q: METTL5‑mediated m6A modification of 18S rRNA drives oral squamous cell carcinoma progression by enhancing CCND3 translation. Oncol Lett 31: 137, 2026.
APA
Liu, L., Sheng, S., Zheng, X., Huang, S., Wang, L., Shu, D. ... He, Q. (2026). METTL5‑mediated m6A modification of 18S rRNA drives oral squamous cell carcinoma progression by enhancing CCND3 translation. Oncology Letters, 31, 137. https://doi.org/10.3892/ol.2026.15490
MLA
Liu, L., Sheng, S., Zheng, X., Huang, S., Wang, L., Shu, D., Tang, L., He, Q."METTL5‑mediated m6A modification of 18S rRNA drives oral squamous cell carcinoma progression by enhancing CCND3 translation". Oncology Letters 31.4 (2026): 137.
Chicago
Liu, L., Sheng, S., Zheng, X., Huang, S., Wang, L., Shu, D., Tang, L., He, Q."METTL5‑mediated m6A modification of 18S rRNA drives oral squamous cell carcinoma progression by enhancing CCND3 translation". Oncology Letters 31, no. 4 (2026): 137. https://doi.org/10.3892/ol.2026.15490
Copy and paste a formatted citation
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Spandidos Publications style
Liu L, Sheng S, Zheng X, Huang S, Wang L, Shu D, Tang L and He Q: METTL5‑mediated m6A modification of 18S rRNA drives oral squamous cell carcinoma progression by enhancing CCND3 translation. Oncol Lett 31: 137, 2026.
APA
Liu, L., Sheng, S., Zheng, X., Huang, S., Wang, L., Shu, D. ... He, Q. (2026). METTL5‑mediated m6A modification of 18S rRNA drives oral squamous cell carcinoma progression by enhancing CCND3 translation. Oncology Letters, 31, 137. https://doi.org/10.3892/ol.2026.15490
MLA
Liu, L., Sheng, S., Zheng, X., Huang, S., Wang, L., Shu, D., Tang, L., He, Q."METTL5‑mediated m6A modification of 18S rRNA drives oral squamous cell carcinoma progression by enhancing CCND3 translation". Oncology Letters 31.4 (2026): 137.
Chicago
Liu, L., Sheng, S., Zheng, X., Huang, S., Wang, L., Shu, D., Tang, L., He, Q."METTL5‑mediated m6A modification of 18S rRNA drives oral squamous cell carcinoma progression by enhancing CCND3 translation". Oncology Letters 31, no. 4 (2026): 137. https://doi.org/10.3892/ol.2026.15490
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