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YTHDC2 inhibits HPV‑positive cervical cancer growth by suppressing SLC7A11 in a ferroptosis‑dependent manner

  • Authors:
    • Li Ren
    • Junjun Zhang
    • Jinjin Yang
    • Li Ji
    • Jingjing Guo
  • View Affiliations / Copyright

    Affiliations: Department of Gynecology, The First Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China, Department of Gynecology, The First Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China
    Copyright: © Ren et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 498
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    Published online on: August 27, 2025
       https://doi.org/10.3892/ol.2025.15244
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Abstract

Chronic infection with high‑risk human papillomavirus (HPV) types increases the risk of developing cervical cancer (CC). Notably, these HPV types are implicated in ~70% of all CC cases. YTH N6‑methyladenosine RNA‑binding protein C2 (YTHDC2) is an N6‑methyladenosine reader associated with several cancers, although its specific function in CC remains poorly understood. The present study aimed to elucidate new functions of YTHDC2 in HPV‑positive CC cell proliferation and ferroptosis, mediated by pathways dependent on protein synthesis. A comparative analysis revealed reduced levels of YTHDC2 in HPV‑positive CC samples and cell lines compared with those in HPV‑negative samples. YTHDC2 overexpression diminished cell viability and proliferation, whereas silencing YTHDC2 enhanced these processes, according to Cell Counting Kit‑8 and colony formation assays. Cells overexpressing YTHDC2 exhibited characteristics of ferroptosis, such as apoptosis, reactive oxygen species production and imbalances in several ferroptosis indicators, including solute carrier family 7 member 11 (SLC7A11). A bioluminescent assay using a firefly luciferase construct fused with the SLC7A11 5'‑untranslated region (UTR) demonstrated a reduction in luminescence in cells exhibiting enhanced YTHDC2 expression. This indicated that YTHDC2 catalyzes translation initiation via deconvolution of the 5'‑UTR of SLC7A11 mRNA. Alterations to the 5'‑UTR led to increased luciferase activity. Furthermore, increased SLC7A11 levels mitigated the effects of YTHDC2 overexpression on proliferation and ferroptosis in CC cells. In conclusion, the results of the present study suggest that low expression of the RNA helicase YTHDC2 contributes to CC proliferation and inhibition of ferroptosis by promoting SLC7A11 translation in HPV‑positive CC cells.
View Figures

Figure 1

Analysis of YTHDC2 expression in
clinical and cell samples of HPV-positive cervical cancer. (A)
RT-qPCR results demonstrating YTHDC2 expression in tissue samples
from HPV-positive (n=25) and HPV-negative (n=25) cervical cancer.
(B) Western blot analysis of the expression of YTHDC2 in tissue
samples from HPV-positive (n=3) and HPV-negative (n=3) cervical
cancer from the aforementioned 25 HPV-positive tissues and 25
HPV-negative CC samples. (C) Immunohistochemistry assessment of the
expression of YTHDC2 in the tissue samples from HPV-positive and
HPV-negative cervical cancer. Magnification, ×40. Analysis of
YTHDC2 mRNA and protein levels in cervical cancer cell lines
(CaSKi, HFF-1, SiHa, SW756, HeLa, DoTc2 4510, C33A and H8) using
(D) RT-qPCR and (E) western blotting, respectively. *P<0.05;
**P<0.01. YTHDC2, YTH N6-methyladenosine RNA-binding protein C2;
HPV, human papillomavirus; RT-qPCR, reverse
transcription-quantitative PCR.

Figure 2

Modulation of YTHDC2 expression in
SiHa and CaSKi cells. SiHa and CaSKi cells were either untreated or
transfected with pCMV1-empty, pCMV1-YTHDC2, control siRNA or YTHDC2
siRNA for 48 h. Reverse transcription-quantitative PCR of (A) SiHA
and (B) CaSKi cells, and western blotting analyses of (C) SiHa and
(D) CaSKi cells, demonstrating the changes in YTHDC2 expression
levels. **P<0.01. YTHDC2, YTH N6-methyladenosine RNA-binding
protein C2; si, small interfering; OE, overexpression; NTC,
non-transfected control.

Figure 3

Effect of YTHDC2 on cell viability
and proliferation in SiHa and CaSKi cells. SiHa and CaSKi cells
were subjected to either no treatment or were transfected with
pCMV1-empty, pCMV1-YTHDC2, control siRNA or YTHDC2 siRNA for 48 h.
Post-transfection cell viability of (A) SiHA and (B) CaSKi cells
was assessed at 48 h using CCK-8 assays. Colony formation assays
were performed to evaluate the proliferation rates of (C) SiHa and
(D) CaSKi cells. *P<0.05; **P<0.01. YTHDC2, YTH
N6-methyladenosine RNA-binding protein C2; si, small interfering;
OE, overexpression; NTC, non-transfected control.

Figure 4

Effect of YTHDC2 on ferroptosis in
SiHa and CaSKi cells. SiHa and CaSKi cells were either untreated or
transfected with pCMV1-empty, pCMV1-YTHDC2, control siRNA or YTHDC2
siRNA for 48 h. Flow cytometry was performed to assess the
apoptosis rates in (A) SiHA and (B) CaSKi cells. ROS levels were
assessed after a 10 min exposure to the ROS-sensitive fluorescent
dye H2DCF-DA (5 µM) in (C) SiHa and (D) CaSKi cells. Western
blotting was performed to assess the levels of proteins involved in
ferroptosis, including p53, ACSL4, GPX4 and SLC7A11, in (E) SiHa
and (F) CaSKi cells. *P<0.05; **P<0.01. YTHDC2, YTH
N6-methyladenosine RNA-binding protein C2; si, small interfering;
OE, overexpression; NC, negative control; ROS, reactive oxygen
species; ACSL4, acyl-CoA synthetase long chain family member 4;
GPX4, glutathione peroxidase 4; SLC7A11, solute carrier family 7
member 11; NTC, non-transfected control.

Figure 5

Role of YTHDC2 in N6-methyladenosine
methylation and suppression of SLC7A11 mRNA expression. CaSKi cells
were either untreated or transfected with pCMV1-empty,
pCMV1-YTHDC2, control siRNA or YTHDC2 siRNA for 48 h. Reverse
transcription-quantitative PCR of (A) SiHA and (B) CaSKi cells
cells revealed the changes in SLC7A11 expression levels. Luciferase
assays were performed on (C) SiHa and (D) CaSKi cells transfected
with either YTHDC2 overexpression or silencing vectors alongside a
reporter plasmid, including the SLC7A11 5′-UTR associated with
luciferase sequences. Luciferase activity ratios corresponding with
the SLC7A11 5′-UTR were normalized to controls. Additional
luciferase assays involved (E) SiHa and (F) CaSKi cells transfected
with either WT or mutant SLC7A11 5′-UTR luciferase reporters.
*P<0.05; **P<0.01; ***P<0.001. YTHDC2, YTH
N6-methyladenosine RNA-binding protein C2; SLC7A11, solute carrier
family 7 member 11; UTR, untranslated region; si, small
interfering; OE, overexpression; NC, negative control; WT,
wild-type; NTC, non-transfected control.

Figure 6

Augmentation of SLC7A11 expression in
YTHDC2-overexpressing SiHa and CaSKi cells. SiHa and CaSKi cells
were co-transfected with pCMV1-YTHDC2 along with either
pcDNA3.1-empty or pcDNA3.1-SLC7A11 for 48 h. Reverse
transcription-quantitative PCR of (A) SiHa and (B) CaSKi cells, and
western blot analysis of (C) SiHa and (D) CaSKi cells were
performed to assess SLC7A11 mRNA and protein levels, respectively,
demonstrating the effects of co-transfection on these cells.
**P<0.01. SLC7A11, solute carrier family 7 member 11; YTHDC2,
YTH N6-methyladenosine RNA-binding protein C2; OE, overexpression;
NC, negative control.

Figure 7

Mitigating effects of SLC7A11
overexpression on YTHDC2-induced changes in SiHa and CaSKi cell
viability. SiHa and CaSKi cells were subjected to co-transfection
with pCMV1-YTHDC2 and either pcDNA3.1-empty or pcDNA3.1-SLC7A11 for
48 h. (A) Cell viability was assessed at 48 h post-transfection
utilizing CCK-8 assays. (B) Colony formation assays were performed
to assess the proliferation rates of these cells. **P<0.01;
***P<0.001. SLC7A11, solute carrier family 7 member 11; YTHDC2,
YTH N6-methyladenosine RNA-binding protein C2; OE, overexpression;
NC, negative control.

Figure 8

Effect of SLC7A11 on counteracting
YTHDC2-driven ferroptosis in SiHa and CaSKi cells. SiHa and CaSKi
cells were simultaneously transfected with pCMV1-YTHDC2 and either
pcDNA3.1-empty or pcDNA3.1-SLC7A11 for 48 h. Flow cytometry was
used to assess ferroptosis in (A) SiHa and (B) CaSKi cells.
Reactive oxygen species accumulation was assessed in (C) SiHa and
(D) CaSKi cells after treating the cells with the H2DCF-DA
fluorescent marker (5 µM) for 10 min. (E) GPX4, ACSL4, p53 and
SLC7A11 protein levels were analyzed using western blotting.
*P<0.05; **P<0.01. SLC7A11, solute carrier family 7 member
11; YTHDC2, YTH N6-methyladenosine RNA-binding protein C2; ACSL4,
acyl-CoA synthetase long chain family member 4; GPX4, glutathione
peroxidase 4; OE, overexpression; NC, negative control.
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Spandidos Publications style
Ren L, Zhang J, Yang J, Ji L and Guo J: YTHDC2 inhibits HPV‑positive cervical cancer growth by suppressing SLC7A11 in a ferroptosis‑dependent manner. Oncol Lett 30: 498, 2025.
APA
Ren, L., Zhang, J., Yang, J., Ji, L., & Guo, J. (2025). YTHDC2 inhibits HPV‑positive cervical cancer growth by suppressing SLC7A11 in a ferroptosis‑dependent manner. Oncology Letters, 30, 498. https://doi.org/10.3892/ol.2025.15244
MLA
Ren, L., Zhang, J., Yang, J., Ji, L., Guo, J."YTHDC2 inhibits HPV‑positive cervical cancer growth by suppressing SLC7A11 in a ferroptosis‑dependent manner". Oncology Letters 30.5 (2025): 498.
Chicago
Ren, L., Zhang, J., Yang, J., Ji, L., Guo, J."YTHDC2 inhibits HPV‑positive cervical cancer growth by suppressing SLC7A11 in a ferroptosis‑dependent manner". Oncology Letters 30, no. 5 (2025): 498. https://doi.org/10.3892/ol.2025.15244
Copy and paste a formatted citation
x
Spandidos Publications style
Ren L, Zhang J, Yang J, Ji L and Guo J: YTHDC2 inhibits HPV‑positive cervical cancer growth by suppressing SLC7A11 in a ferroptosis‑dependent manner. Oncol Lett 30: 498, 2025.
APA
Ren, L., Zhang, J., Yang, J., Ji, L., & Guo, J. (2025). YTHDC2 inhibits HPV‑positive cervical cancer growth by suppressing SLC7A11 in a ferroptosis‑dependent manner. Oncology Letters, 30, 498. https://doi.org/10.3892/ol.2025.15244
MLA
Ren, L., Zhang, J., Yang, J., Ji, L., Guo, J."YTHDC2 inhibits HPV‑positive cervical cancer growth by suppressing SLC7A11 in a ferroptosis‑dependent manner". Oncology Letters 30.5 (2025): 498.
Chicago
Ren, L., Zhang, J., Yang, J., Ji, L., Guo, J."YTHDC2 inhibits HPV‑positive cervical cancer growth by suppressing SLC7A11 in a ferroptosis‑dependent manner". Oncology Letters 30, no. 5 (2025): 498. https://doi.org/10.3892/ol.2025.15244
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