YTHDF2 Promotes the Metastasis of Oral Squamous Cell Carcinoma through the JAK- STAT Pathway

doi:10.21203/rs.3.rs-5244964/v1

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YTHDF2 Promotes the Metastasis of Oral Squamous Cell Carcinoma through the JAK- STAT Pathway

https://doi.org/10.21203/rs.3.rs-5244964/v1

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Background: RNA-binding proteins act as crucial mediators between m6A modification and RNA function, playing a significant role in the recurrence and metastasis of oral squamous cell carcinoma. YTHDF2, the first identified RNA-binding protein, is associated with the prognosis of certain cancer patients. However, the role of YTHDF2 in oral squamous cell carcinoma and its underlying molecular mechanism have not been fully explored.

Methods: First, we analysed the expression levels of YTHDF2 and itscorrelation with clinical features via the use of public databases and oral squamous cell carcinoma patient samples. Subsequently, in vitro functional experiments were conducted to assess the effects of YTHDF2 on the proliferation, migration and invasion capabilities of oral squamous cell carcinoma cells. Additionally, RNA-seq analysis was utilized to investigate the signalling pathways modulated by YTHDF2, followed by experimental validation.

Results: Our findings revealed that YTHDF2 expression was significantly elevated in oral squamous cell carcinoma tissues and cells, with levels significantly correlated with the clinical stage, pathological grade, and survival time of patients. The knockdown of YTHDF2 resulted in decreased proliferation, migration and invasion abilities in oral squamous cell carcinoma cells. Furthermore, RNA sequencing data indicated that silencing YTHDF2 suppressed the JAK-STAT signalling pathway, and the use of STAT3 activators reversed this suppressive effect in oral squamous cell carcinoma cells.

Conclusion: Our study demonstrated that YTHDF2 promotes the proliferation, metastasis and invasion of oral squamous cell carcinoma by positively regulating the JAK-STAT signalling pathway, suggesting that YTHDF2 could serve as a potential prognostic marker for oral squamous cell carcinoma.

YTHDF2

Oral squamous cell carcinoma

Metastasis

JAK-STAT pathway

m6A

Oral squamous cell carcinoma (OSCC) is one of the most common oral cancers worldwide and is often linked to smoking, alcohol consumption, and human papillomavirus infection.^[1] OSCC accounts for more than 90% of all oral tumours, with over 500,000 new cases and 170,000 fatalities reported annually, with increasing incidence. It is expected that by 2030, the annual incidence rate of OSCC will increase to 30%, resulting in 1.08 million new cases.^[2] Unfortunately, the early diagnosis rate of OSCC is low and often coincides with lymph node metastasis. Approximately 70% of patients are diagnosed at an advanced stage, with obvious lymph node metastasis; this results in an approximate 50% decrease in the 5-year survival rate.^[3] The core reason for this situation is that the main factors contributing to the poor therapeutic effects on OSCC—local recurrence and distant metastasis—have not been effectively resolved.^[4] Therefore, understanding the molecular mechanisms underlying OSCC recurrence and metastasis as well as finding new therapeutic targets are crucial areas of current research.

During the process of OSCC metastasis, cells develop the ability to metastasize through genetic or epigenetic changes. Among these changes, epigenetic modifications at the RNA level, particularly 6-methyladenine (N6-methyladenosing, m6A), have garnered significant attention.^[5] m6A is the most common RNA modification in eukaryotes, and its addition and removal are dynamic and reversible processes.^[6] This process is regulated by methyltransferases, which add m6A; demethylases, which remove m6A; and m6A-binding proteins.^[6] m6A-binding proteins play crucial roles in connecting m6A modification to RNA function. Specific target genes of m6A modification regulate the splicing, processing, transport, translation, and stability of RNA, thereby impacting the proliferation, migration, invasion, and chemotherapy resistance of tumour cells.^[7] Notably, YTHDF2, identified as the first and most effective m6A reading protein, has been found to have dual functions in regulating the proliferation and migration of tumour cells.^[8] In certain types of cancers, such as acute myeloid leukaemia,^[9] lung cancer,^[10] and bladder cancer,^[11] YTHDF2 is upregulated and functions as an oncogene. Conversely, in osteosarcoma^[12] and melanoma,^[13] YTHDF2 is downregulated and acts as a tumour suppressor. Nevertheless, there is little research on the role of YTHDF2 in OSCC.

This study analysed the expression of YTHDF2 in OSCC tissues compared with that in control tissues, as well as the relationships between YTHDF2 levels and the clinical characteristics and survival times of OSCC patients. Functional experiments were conducted to elucidate the regulatory effect of YTHDF2 on OSCC proliferation and metastasis. Additionally, transcriptome sequencing was performed to identify the signalling pathways through which YTHDF2 regulates the biological behaviour of OSCC cells. Overall, this research provides insights into the molecular mechanisms underlying YTHDF2-mediated proliferation and metastasis in OSCC, suggesting a potential therapeutic target for future studies.

Clinical samples

The study analysed a group of 59 squamous cell carcinoma tissue samples and 20 paracancerous tissue samples from patients treated at the Department of Oral and Maxillofacial Surgery, Stomatological Hospital Affiliated with Southwest Medical University, between January 2018 and December 2020. The follow-up deadline was January 2024. None of the patients had undergone preoperative radiation or chemotherapy, and all tissue samples were obtained with appropriate informed consent and approval from the Institutional Review Board of the Stomatological Hospital Affiliated with Southwest Medical University.

IHC staining

All the tissue sections were deparaffinized, rehydrated, subjected to antigen retrieval, blocked and incubated with an anti-YTHDF2 antibody (1:2000, Abcam) overnight at 4°C. This was followed by incubation with secondary antibodies and detection via a diaminobenzidine (DAB) kit (Gene Tech, China), followed by haematoxylin counterstaining. The images were captured via an upright microscope system (Nikon, Japan), and the YTHDF2 immunostaining score was calculated by multiplying the score for the proportion of positively stained tumour cells (categories: 1 (< 25%), 2 (25–50%), 3 (50–75%), and 4 (> 75%) by the score for staining intensity (categories: 1-weak staining, 2-moderate staining, and 3-strong staining). The staining score was assessed by two blinded pathologists, and patients were categorized into low-staining (1–6) and high-staining (8–12) groups. Detailed patient information can be found in Table S1.

Cell culture and transfection

SCC9 and NOK cells were kindly provided by Professor Wang.^[14] SCC9 cells were cultured in DMEM/F12 (Gibco, USA), and NOK cells were cultured in high-glucose DMEM (Gibco, USA). Both growth media were supplemented with 10% foetal bovine serum (FBS) (Gibco, USA) and 1% penicillin and streptomycin (Beyotime Biotech, China). The cells were kept in a 5% carbon dioxide incubator, with regular medium changed 2–3 times every week. The cells were passaged when they reached 80% confluence.

Plasmid transfection was conducted via a siRNA transfection kit (Ribo, China) following the manufacturer’s instructions, with three YTHDF2 siRNA sequences designed: siYTHDF2-1, GACCAAGAATGGCATTGCA; siYTHDF2-2, GCACAGAAGTTGCAAGCAA; and siYTHDF2-3, GGTAGCGGGTCCATTACTA.

Quantitative real-time PCR (qRT‒PCR)

Total RNA was extracted from cells via the Steady Pure Rapid RNA Extraction Kit according to the manufacturer’s instructions. This was followed by cDNA synthesis with ReverTra Ace qPCR RT Master Mix (Toyobo, Japan). PCR was then conducted via the SYBR Green Master Mix Kit (Takara, Japan) on a StepOne Plus Real-time PCR System (Applied Biosystems, USA), with GAPDH as the internal control. Each sample was analysed in triplicate, with data analysis based on the 2^−ΔΔCT method. The primers used are listed in Table S2.

Western blot

Cellular proteins were extracted by lysing cells in RIPA buffer supplemented with protease inhibitors (Sigmar, USA). The extracted protein concentrations were detected via a BCA kit. Subsequently, 20 µg of protein per sample was separated via 10% sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE) and transferred onto a PVDF membrane (Millipore, USA). The samples were incubated overnight at 4°C with primary antibodies against YTHDF2 (1:1000, Abcam), JAK2 (1:5000, Abcam), STAT3 (1:1000, Abcam), p-JAK2 (1:5000, Abcam), p-STAT3 (1:1000, Abcam) and GAPDH (1:10000, affinity), followed by the application of an HRP-labelled secondary antibody (1:1000, Beyotime). The protein‒antibody complexes were visualized with enhanced chemiluminescence (ECL) detection reagents and analysed via Image Lab software.

Proliferative assay

Cell proliferation was assessed through CCK-8 and colony formation assays. For the CCK-8 assay, cells were plated in 96-well plates at a density of 2000 cells per well. The absorbance was measured at 0, 24, 48, 72, and 96 hours at a wavelength of 450 nm postseeding via the CCK-8 system (Dojindo, Japan). For the colony formation assay, cells were seeded in 6-well plates at a density of 1000 cells per well and cultured in DMEM/F12 medium supplemented with 10% FBS for approximately 2 weeks. Colonies were visualized and quantified after fixation with methanol and 0.1% crystal violet staining for 30 minutes.

Scratch wound assay

The cells were seeded into 6-well plates and allowed to grow until they reached confluence. A scratch of approximately 1 mm in the confluent cell layer was made via a P200 pipette tip, followed by continued cell culture. Images were captured at 0, 24, 48, and 72 hours, and the reduction in the wound area was analysed via ImageJ software.

Cell migration and invasion assays

Cell migration and invasion abilities were assessed using chambers with or without Matrigel. A cell suspension was prepared in serum-free DMEM/F12 medium at a concentration of 1🞨10⁵; 500 µL of the suspension was placed in the upper chamber; and 1 ml of DMEM/F12 medium containing 10% FBS was added to the lower chamber. After incubation for 24 hours, the cells on the upper surface of the filter were removed, and those that had migrated through the filter and adhered to the lower surface were stained with 0.1% crustal violet. Five random fields of view were selected under a microscope, images were captured, and the cells were quantified.

Transcriptome sequencing

Total RNA was extracted from three groups of SCC9 cells transfected with siYTHDF2 or control plasmid and sent to Lianchuan Biotech Biotechnology Co., Ltd. (HangZhou, China) for transcriptome sequencing. The RNA was initially tested for quantity, purity, and integrity. cDNAoligo(dT) magnetic beads and high-temperature fragmentation technology were subsequently used to capture and fragment mRNAs with a PolyA tail. This was followed by reverse transcription for conversion into cDNA and second-strand synthesis, with a fragment size of approximately 300 bp ± 50 bp, for library construction. Paired-end sequencing was then conducted via an Illumina Novaseq™ 6000 in PE150 mode. The sequencing data underwent quality control procedures, including the removal of adapters and the filtering of low-quality and repetitive sequences. Alignment and transcript assembly were performed via HISAT2 and StringTie software programs. FPKM quantification was carried out via the ballgown package, and significant differences between samples were analysed via the R package edgeR. Genes with a fold change (FC) greater than 2 or less than 0.5 and a p value less than 0.05 were considered differential genes, confirmed by GO and KEGG enrichment analysis.

Bioinformatic analysis

Pancancer RNA expression data were obtained from the TCGA (https://portal.gdc.cancer.gov/) database in three standardized forms: TPM, FPKM, and RPM. OSCC data were extracted from various oral cancer sites within TCGA-HNSC, consisting of 362 OSCC samples and 32 adjacent cancer samples. Clinical information and RNA-seq expression profiles were downloaded for subsequent analysis.

The “limma” package in R (version 4.0.3) was utilized for data integration and analysis, with a focus on differentially expressed genes, through extracting gene names, p values, false discovery rates (FDRs), and log2-fold change values. Genes with incomplete information were excluded, and those meeting the criteria of FDR < 0.05 and |1log2FC| > 3 were subjected to further analysis. Differential gene visualization was carried out via the “ggplot 2” package. Kaplan‒Meier overall survival curves were constructed with the “survial” and “survminer” packages, while ROC analysis was conducted with the “pROC” package, and ROC curves were visualized via “ggplot2”. The area under the ROC curve served as an indicator of accuracy in diagnosis and prognosis prediction.

Statistical analysis

Data analysis and statistics were conducted via R and SPSS 23.0. Student's t test was utilized to compare two subgroups, whereas one-way ANOVA was used for comparisons among more than two subgroups. Kaplan‒Meier survival analysis was conducted to assess the differences in overall survival (OS) based on different YTHDF2 expression levels. A p value less than 0.05 was considered statistically significant.

Upregulated expression of YTHDF2 is correlated with poor prognosis in oral squamous cell carcinoma

To investigate YTHDF2 expression in different cancer types, we analysed the YTHDF2 levels in 33 cancer tissues and compared them to normal tissues. YTHDF2 was found to be upregulated in 13 cancer types, including bladder urothelial carcinoma (BLCA), breast invasive carcinoma (BRCA), cervical squamous cell carcinoma, endocervical adenocarcinoma (CESC), cholangiocarcinoma (CHOL), colon adenocarcinoma (COAD), oesophageal carcinoma (ESCA), glioblastoma multiforme (GBM), head and neck squamous cell carcinoma (HNSC), liver hepatocellular carcinoma (LIHC), lung adenocarcinoma (LUAD), prostate adenocarcinoma (PRAD), stomach adenocarcinoma (STAD), and uterine corpus endometrial carcinoma (UCEC) (Fig. 1A).

In the context of OSCC, significantly higher YTHDF2 expression was observed in OSCC tissues than in normal tissues (Fig. 1B). Higher YTHDF2 expression levels were significantly correlated with advanced tumour grade, as shown in Fig. 1C. OSCC patients were divided into high and low YTHDF2 expression groups on the basis of average YTHDF2 levels, and a Kaplan‒Meier curve revealed that the survival time of the high YTHDF2 expression group was significantly lower than that of the low-expression group (Fig. 1D). Furthermore, the diagnostic potential of YTHDF2 expression in OSCC was assessed via receiver operating characteristic (ROC) curve analysis. As shown in Fig. 1E, the area under the curve (AUC) for YTHDF2 was 0.748, with a 95% confidence interval (CI) ranging from 0.664–0.831.

Figure 1. Upregulated expression of YTHDF2 correlates with poor prognosis in oral squamous cell carcinoma. (A) The expression levels of YTHDF2 in various cancers; (B) Statistical analysis of YTHDF2 levels in OSCC tissues and normal oral mucosa tissues; (C) Statistical analysis of YTHDF2 levels in different clinical grades; (D) Kaplan‒Meier survival curve based on YTHDF2 expression levels; (E) ROC curve analysis of the potential diagnostic value of the YTHDF2 expression level in OSCC diagnosis.

YTHDF2 upregulation was associated with poor prognosis in OSCC

Immunohistochemical staining was conducted on clinical tissue samples to further investigate the correlation between YTHDF2 and the clinical characteristics of OSCC. The results revealed a predominant cytoplasmic expression of YTHDF2 in OSCC tissues through a higher percentage of positive staining and a more intense staining colour (Fig. 2A). Statistical analysis revealed significantly higher YTHDF2 staining scores in OSCC tissues than in control tissues (Fig. 2B). Furthermore, higher levels of YTHDF2 expression were correlated with advanced tumour stage (T_3 + 4 and T_1 + 2), advanced clinical stage (C_III+IV and C_I+II), and lymph node metastasis (L_N+ vs. L_N-) (Fig. 2B). K‒M analysis revealed a significantly shorter overall survival time in patients with high YTHDF2 expression than in those with low YTHDF2 expression (Fig. 2C). The area under the ROC curve was 0.733 (95% CI: 0.626–0.841) (Fig. 2D), indicating that the level of YTHDF2 expression could be a valuable diagnostic marker for OSCC. A staining score cut-off value of 6 was identified for distinguishing OSCC patients from healthy controls.

Figure 2. YTHDF2 was upregulated and associated with poor prognosis in OSCC. (A) Results of IHC staining for YTHDF2 in OSCC tissues and paracancerous tissues. (B) Statistical analysis of the staining scores of YTHDF2 in OSCC tissues and paracancerous tissues and the relationships between YTHDF2 expression levels and tumour stage (T3 + 4 vs. T1 + 2), clinical stage (CIII + IV vs. CI + II) and lymph node metastasis (LN + vs. LN−). (C) Kaplan‒Meier survival curve based on YTHDF2 expression levels. (D) ROC curve of the YTHDF2 expression level in OSCC.

Knockdown of YTHDF2 suppresses OSCC cell proliferation, migration and invasion

To investigate the impact of YTHDF2 on OSCC cell function, three siRNAs targeting YTHDF2 were synthesized and transfected into SCC9 cells. The knockdown efficiency of these siRNAs in SCC9 cells was determined via RT‒PCR and Western blotting (Fig. 3A, 3B). Among the tested siRNAs, siRNA-1 showed the most effective silencing and was selected for further experiments. Cell proliferation was assessed through CCK-8 and colony formation assays, which revealed that YTHDF2 knockdown significantly inhibited the growth of SCC9 cells (Fig. 3C, 3D). Additionally, transwell migration and invasion assays indicated that YTHDF2 knockdown resulted in decreased migration and invasion capabilities of SCC9 cells (Fig. 3E, 3F). The scratch wound assay further confirmed the decreased migration ability of SCC9 cells following YTHDF2 silencing (Fig. 3G, 3H).

Fig. 3. Knockdown of YTHDF2 suppressed OSCC cell proliferation, migration and invasion. The knockdown of YTHDF2 was confirmed through (A) real-time quantitative PCR and (B) western blotting. (C) CCK-8 detection results, (D) colony formation analysis, (E-F) migration and invasion results, and (G-H) 24-hour scratch migration results of the YTHDF2-silenced group and the control group.

The JAK-STAT pathway may be a potential mechanism for YTHDF2-regulated progression of OSCC

To investigate the potential mechanism underlying the oncogenic role of YTHDF2 in OSCC, we conducted transcriptome sequencing on YTHDF2-silenced cells and control cells. The results revealed a total of 1621 differentially expressed genes in cells after YTHDF2 knockdown, including 740 downregulated genes and 881 upregulated genes (Fig. 4A). Among the thousands of potential targets, STAT1 and STAT2 were significantly upregulated during YTHDF2 knockdown (Fig. 4B). Furthermore, GO analysis of the differentially expressed genes highlighted the critical involvement of YTHDF2 in key processes such as cell proliferation, the inflammatory response, apoptosis, and the immune response in OSCC (Fig. 4C). KEGG analysis further revealed notable alterations in pathways, including the JAK-STAT signalling, Toll-like receptor signalling, NOD-like receptor signalling, P53 signalling, and TNF signalling pathways, following YTHDF2 knockdown (Fig. 4D). Additionally, increased expression levels of STAT3 were observed in OSCC tissues compared with normal tissues (Fig. 4E), with a positive correlation between YTHDF2 expression and JAK2 and STAT3 expression (Fig. 4F). These findings suggest that the JAK-STAT signalling pathway could be a potential mechanism by which YTHDF2 regulates OSCC behaviour.

Figure 4. YTHDF2 regulates the JAK-STAT signalling pathway. (A) Statistical analysis of the upregulated or downregulated genes following the knockdown of YTHDF2. (B) Volcano plot of the differentially expressed genes after the knockdown of YTHDF2. (C) GO and (D) KEGG analyses of the possible signalling pathways regulated by YTHDF2. (E) JAK2 and STAT3 expression levels in OSCC tissues and normal tissues. (F) Correlations between JAK2, STAT3 and YTHDF2 expression in OSCC.

YTHDF2 promoted OSCC growth and metastasis via the JAK-STAT pathway

To confirm that YTHDF2 regulates cell behaviour via the JAK-STAT pathway, we first analysed the protein expression of JAK2 and STAT3 in SCC9 cells after YTHDF2 knockdown. Our results revealed no change in nonphosphorylated JAK2 or STAT3 levels but did reveal a significant decrease in the levels of phosphorylated JAK2 and STAT3 (Fig. 5A, 5B). In a rescue experiment, YTHDF2-silenced SCC9 cells were subsequently treated with the STAT3 activator Colivelin. Cell proliferation was assessed via CCK-8 and colony formation assays, whereas cell migration and invasion were analysed via transwell migration, scratch, and invasion assays. As shown in Fig. 5C-5H, the cells treated with Colivelin exhibited increased proliferation, migration, and invasion capabilities compared with those in the YTHDF2-silenced group.

Fig. 5. YTHDF2 promoted OSCC growth and metastasis via the JAK-STAT pathway. (A) Expression levels and (B) statistical analysis of the JAK and STAT3 proteins between the YTHDF2-silenced group and the control group. (C) CCK-8 assay results, (D) colony formation analysis, (E-F) migration and invasion results, and (G-H) 24-h scratch migration results of the YTHDF2-silenced cells with or without treatment with the STAT3 activator colivelin.

This study revealed that YTHDF2 is significantly overexpressed in OSCC and is closely associated with the clinical stage, grade, and survival time of OSCC patients. Further investigation into the impact of YTHDF2 on OSCC cell behaviour revealed that downregulation of YTHDF2 led to a decrease in the proliferation, migration, and invasion capabilities of OSCC cells, supporting its role in promoting cancer cell growth and invasion. Transcriptome sequencing results indicated that YTHDF2 may modulate the JAK-STAT signalling pathway. The activation of the JAK-STAT pathway in YTHDF2-knockdown cells partially restored their inhibited proliferation, migration, and invasion abilities, suggesting that YTHDF2 promotes the progression of OSCC through the JAK-STAT signalling pathway.

YTHDF2 primarily regulates mRNA degradation, and its role in various tumours is still debated.^[15] While most studies have shown high expression of YTHDF2 in pancreatic cancer,^[16] liver cancer,^[17] ovarian cancer,^[18] cervical cancer,^[19] as well as promoting tumour progression; a minority of scholars have reported a tumour suppressor role for YTHDF2 in certain cancers. For example, Hou^[20] et al. reported significant downregulation of YTHDF2 in hepatocellular carcinoma cells, leading to inflammation, vascular abnormalities, and metastasis. Similarly, Shen^[21] et al. reported a suppressive function of YTHDF2 in gastric cancer through the destabilization of FOXC2 mRNA. This discrepancy may be attributed to YTHDF2 targeting different mRNAs, including tumour suppressor genes and oncogenes. Although research on YTHDF2 and OSCC is limited, our study revealed that high expression of YTHDF2 in OSCC was correlated with advanced clinical stages, high pathological grades, and short survival times. These results underscore the oncogenic role of YTHDF2 in OSCC and contribute to filling a gap in the current literature.

The JAK-STAT signalling pathway is crucial for regulating various cellular functions, such as proliferation, migration, differentiation, and apoptosis, and plays a significant role in tumour development.^[22][23] Activation of this pathway involves JAK initiating tyrosine phosphorylation of receptors thereby recruiting STATs, which form dimers and enter the nucleus to regulate gene transcription.^[24][25] Among the STAT family members, STAT3 is particularly important in transmitting signals from the cell membrane to the nucleus,^[26] and its abnormal expression is associated with multiple cancers, including breast,^[27] ovarian,^[28] and lung cancers.^[29] Our research confirmed the high expression of STAT3 in OSCC and revealed the role of YTHDF2 in activating the JAK-STAT pathway to promote OSCC cell malignancy. Despite the tumour-promoting effects of JAK-STAT signalling, some preclinical studies have shown that JAK inhibitors may compromise NK cell antitumour immunity and enhance breast cancer metastasis.^[30] This discrepancy could be due to specific JAK ligands being targeted. In our future research, we aim to further understand how YTHDF2 modulates the JAK-STAT pathway, potentially providing new insights for targeted therapeutic strategies.

In summary, our study demonstrated the critical function of YTHDF2 in OSCC, which promotes OSCC proliferation and metastasis by regulating the JAK-STAT signalling pathway.

Acknowledgements

We thank Prof. Anxun Wang (Sun Yat-sen University) for the generous gift of SCC9 and NOK cells.

Authors’ contributions

Zhezheng Chen performed the main parts of the experiment; Dan Zhao analyzed the experimental data and wrote the manuscript; Yamin Yuan, Lu Zeng and Zhengzhou Luo provided technical assistance with the experiments; Junliang Chen, Xiaorong Lan, Yun He and Lin Liu designed and supervised the study. All authors read and approved the final manuscript.

Funding

This work was funded by Luzhou Science and Technology Program (2022-SYF-58), Project of Southwest Medical University (21YYJC0402), Youth Science Climbing Program of The Affiliated Stomatological Hospital of Southwest Medical University (2020QY03) and Sichuan Science and Technology Program (2022YFS0634).

Availability of data and materials

All data can be obtained upon a reasonable request to corresponding authors. TCGA databases are publicly available at(https://portal.gdc.cancer.gov/), respectively.

Ethics approval and consent to participate

All tissue samples were obtained with the appropriate informed consent and approval from the Institutional Review Board of the Stomatological Hospital Affiliated with Southwest Medical University.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

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No competing interests reported.

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YTHDF2 Promotes the Metastasis of Oral Squamous Cell Carcinoma through the JAK- STAT Pathway

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Abstract

Figures

Introduction

Materials and Methods

Clinical samples

IHC staining

Cell culture and transfection

Quantitative real-time PCR (qRT‒PCR)

Western blot

Proliferative assay

Scratch wound assay

Cell migration and invasion assays

Transcriptome sequencing

Bioinformatic analysis

Statistical analysis

Results

Upregulated expression of YTHDF2 is correlated with poor prognosis in oral squamous cell carcinoma

YTHDF2 upregulation was associated with poor prognosis in OSCC

Knockdown of YTHDF2 suppresses OSCC cell proliferation, migration and invasion

The JAK-STAT pathway may be a potential mechanism for YTHDF2-regulated progression of OSCC

YTHDF2 promoted OSCC growth and metastasis via the JAK-STAT pathway

Discussion

Conclusion

Declarations

References

Additional Declarations

Supplementary Files

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