Progression Of Oral Squamous Cell Carcinoma Research Articles

E-cadherin Expression in Premalignant Lesions, Premalignant Conditions, Oral Squamous Cell Carcinoma, and Normal Mucosa: An Immunohistochemical Study.

Background Oral squamous cell carcinoma (OSCC) is a multi-step process. Epithelial-mesenchymal transition (EMT) is an important step in the progression of OSCC. One of the components that influence EMT is E-cadherin. The aim of this study was to determine the expression of E-cadherin in oral submucous fibrosis (OSMF), various grades of epithelial dysplasia, OSCC, and to compare it with the expression in the normal mucosa. Material and methods E-cadherin immunohistochemical detectionwas done using a monoclonal antibody of clone EP-6TM and the PolyExcel HRP/DAB chromogen detection system. A total of 100 samples, were divided into four groups, which included epithelial dysplasia(group 2) (30 cases), oral submucous fibrosis (group 3) (OSMF-30 cases), and oral squamous cell carcinoma (group 4) (OSCC-30 cases), which was compared with normal mucosa (group 1) (10 cases). The positive control used for E-cadherin was ductal breast carcinoma. Results All the cases of normal mucosa, epithelial dysplasia, and OSMF showed positivity for E-cadherin expression. In OSCC, 97% of cases expressed E-cadherin except one case. Out of 30 cases of epithelial dysplasia, 53% of mild epithelial dysplasia had a moderate intensity of expression and 75% had a mild intensity of E-cadherin expression. In moderately differentiated OSCC, 82% of cases showed mild intensity. Tissue localization of the E-cadherin stain in the basal layer decreased from normal mucosa to grades of epithelial dysplasia and OSCC. The pattern of E-cadherin staining in all the cases of group I, group II, and group III was membranous. In 97% of OSCC cases, both membranous and cytoplasmic staining were seen. Conclusion E-cadherin expression was reduced in increasing grades of epithelial dysplasia, OSCC, and OSMF compared to that of normal mucosa. E-cadherin expression is reduced as the lesions progress to malignancy. Hence, E-cadherin can be considereda surrogate marker of malignancy.

THBS1-Mediated Degradation of Collagen via the PI3K/AKT Pathway Facilitates the Metastasis and Poor Prognosis of OSCC.

Oral squamous cell carcinoma (OSCC) is a prevalent form of malignant tumor, characterized by a persistently high incidence and mortality rate. The extracellular matrix (ECM) plays a crucial role in the initiation, progression, and diverse biological behaviors of OSCC, facilitated by mechanisms such as providing structural support, promoting cell migration and invasion, regulating cell morphology, and modulating signal transduction. This study investigated the involvement of ECM-related genes, particularly THBS1, in the prognosis and cellular behavior of OSCC. The analysis of ECM-related gene data from OSCC samples identified 165 differentially expressed genes forming two clusters with distinct prognostic outcomes. Seventeen ECM-related genes showed a significant correlation with survival. Experimental methods were employed to demonstrate the impact of THBS1 on proliferation, migration, invasion, and ECM degradation in OSCC cells. A risk-prediction model utilizing four differentially prognostic genes demonstrated significant predictive value in overall survival. THBS1 exhibited enrichment of the PI3K/AKT pathway, indicating its potential role in modulating OSCC. In conclusion, this study observed and verified that ECM-related genes, particularly THBS1, have the potential to influence the prognosis, biological behavior, and immunotherapy of OSCC. These findings hold significant implications for enhancing survival outcomes and providing guidance for precise treatment of OSCC.

Identification of an endoplasmic reticulum stress-related prognostic risk model with excellent prognostic and clinical value in oral squamous cell carcinoma.

Recently, endoplasmic reticulum stress related gene (ERS) markers have performed very well in predicting the prognosis of tumor patients. The differentially expressed genes in Oral squamous cell carcinoma (OSCC) were obtained from TCGA and GTEx database. Three prognosis-related and differentially expressed ERSs were screened out by Least Absolute Selection and Shrinkage Operator (Lasso) regression to construct a prognostic risk model. Receiver Operating Characteristic Curve (ROC), riskplots and survival curves were used to verify the model's accuracy in predicting prognosis. Multi-omics analysis of immune infiltration, gene mutation, and stem cell characteristics were performed to explore the possible mechanism of OSCC. Finally, we discussed the model's clinical application value from the perspective of drug sensitivity. Three genes used in the model (IBSP, RDM1, RBP4) were identified as prognostic risk factors. Bioinformatics analysis, tissue and cell experiments have fully verified the abnormal expression of these three genes in OSCC. Multiple validation methods and internal and external datasets confirmed the model's excellent performance in predicting and discriminating prognosis. Cox regression analysis identified risk score as an independent predictor of prognosis. Multi-omics analysis found strong correlations between risk scores and immune cells, cell stemness index, and tumor mutational burden (TMB). It was also observed that the risk score was closely related to the half maximal inhibitory concentration of docetaxel, gefitinib and erlotinib. The excellent performance of the nomogram has been verified by various means. A prognostic model with high clinical application value was constructed. Immune cells, cellular stemness, and TMB may be involved in the progression of OSCC.

Overexpression of RAB27A in Oral Squamous Cell Carcinoma Promotes Tumor Migration and Invasion via Modulation of EGFR Membrane Stability.

Oral squamous cell carcinoma (OSCC) is the most prevalent subtype of head and neck tumors, highly prone to lymph node metastasis. This study aims to examine the expression pattern of Ras-related protein Rab-27A (RAB27A) and explore its potential implications in OSCC. The expression of RAB27A was assessed through immunohistochemical analysis utilizing tissue microarrays. In vitro experiments were conducted using RAB27A-knockdown cells to investigate its impact on OSCC tumor cells. Additionally, transcriptome sequencing was performed to elucidate potential underlying mechanisms. RAB27A was significantly overexpressed in OSCC, and particularly in metastatic lymph nodes. It was positively correlated with the clinical progression and poor survival prognosis. Silencing RAB27A notably decreased the proliferation, migration, and invasion abilities of OSCC cells in vitro. A Gene Ontology (GO) enrichment analysis indicated a strong association between RAB27A and the epidermal growth factor receptor (EGFR) signaling pathway. Further investigations revealed that RAB27A regulated the palmitoylation of EGFR via zinc finger DHHC-type containing 13 (ZDHHC13). These findings provide insights into OSCC progression and highlight RAB27A as a potential therapeutic target for combating this aggressive cancer.

Simvastatin inhibits proliferation and promotes apoptosis of oral squamous cell carcinoma through KLF2 signal

ObjectivesThis study aimed to explore the role and specific mechanism of the cholesterol-lowering drug simvastatin in inhibiting oral squamous cell carcinoma (OSCC). MethodsThe proliferation, apoptosis, and migration levels of OSCC cells were detected by CCK8, quantitative real-time polymerase chain reaction, Western blot, colony formation, TdT-mediated dUTP Nick-End Labeling assay, and wound healing assay. The inhibitory effect of simvastatin in vivo was detected by a mouse xenograft tumor model. Immunohistochemistry and immunofluorescence staining were used to assess the KLF2 and β-catenin expressions in cells and tissues. ResultsKLF2 expression in OSCC cells and tissues was downregulated. The addition of KLF2 inducer, GGTI298, inhibited the proliferation and migration of OSCC cells. Simvastatin played a role in inhibiting the proliferation and promoting the apoptosis of OSCC cells. Moreover, it inhibited β-catenin expression and promoted KLF2 expression in OSCC cells. KLF2 siRNA reversed the effect of simvastatin on the proliferation and apoptosis of OSCC cells. ConclusionsKLF2, as a tumor suppressor gene, may be an important marker for diagnosing and treating OSCC. Simvastatin inhibits the progression of OSCC by regulating the KLF2 signal.

TRIM32 promotes oral squamous cell carcinoma progression by enhancing FBP2 ubiquitination and degradation

The aberrant expression of TRIM32, an E3 ubiquitin ligase, has been identified in multiple malignant cancer types. Nevertheless, the functional roles and detailed mechanisms of TRIM32 in oral squamous cell carcinoma (OSCC) remain to be elucidated. Here, we investigated TRIM32 expression and its functional role in OSCC. TRIM32 expression was consistently elevated in OSCC tissues, particularly in samples from patients with advanced clinical grades. Functionally, silencing TRIM32 dampened OSCC cell growth, migration and invasion. Additionally, a xenograft tumor model suggested that TRIM32 knockdown suppressed in vivo OSCC tumor growth and lung metastasis formation. Mechanistically, we discovered that TRIM32 directly bound to the FBP2 protein via mass spectrometry and co-immunoprecipitation. TRIM32 could interact with FBP2 and accelerates its degradation, eventually enhancing glycolysis in OSCC cell lines. Importantly, rescue assays demonstrated that FBP2 silencing could at least partially offset the tumor-suppressive and aerobic glycolysis inhibition effect induced by TRIM32 knockdown. Thus, our findings demonstrate that TRIM32 plays a crucial role in promoting tumor growth and enhancing glycolysis through FBP2 inhibition. Given OSCC is associated with increased glycolysis levels, our study suggests potential therapeutic targets for OSCC treatment.

CircLDLRAD3 inhibits Oral squamous cell carcinoma progression by regulating miR-558/Smad4/TGF-β.

Oral squamous cell carcinoma (OSCC) is a malignant neoplasm with high mortality and morbidity. The role of circRNA and its molecular mechanism in OSCC remains largely unknown. The study aims to explore the role of a novel circular RNA (circLDLRAD3) in OSCC and its underlying mechanism. PCR and fluorescence in situ hybridization were used to explore the expression features of circLDLRAD3 in OSCC. The effects of circLDLRAD3 on the behaviour of OSCC were investigated using CCK-8, colony formation assay, transwell and animal experiments. Bioinformatics analysis along with dual luciferase reporter assay and RIP assay were used to reveal the interaction between circLDLRAD3, miR-558 and Smad4. It was revealed that circLDLRAD3 exhibited low expression status in OSCC. CircLDLRAD3 inhibits proliferation, migration, and invasion of OSCC cells both in vitro and in vivo. Mechanistically, circLDLRAD3 could bind with miR-558 to positively regulate its target gene Smad4 expression. Rescue experiments further confirmed both miR-558 overexpression and Smad4 knockdown could reverse the influence of circLDLRAD3 on OSCC phenotypes. Moreover, circLDLRAD3 regulate the TGF-β signalling pathways to influence EMT through miR-558/Smad4 axis. Our study found that circLDLRAD3 is downregulated in OSCC and verified its tumour suppressor function and mechanism in OSCC through sponging miR-558 to regulate miR-558/Smad4/TGF-β axis. The characterization of such regulating network uncovers an important mechanism underlying OSCC progression, which could provide promising targets targeted therapy strategies for OSCC in the future.

Role of Stromal Fibroblast–Induced WNT7A Associated with Cancer Cell Migration Through the AKT/CLDN1 Signaling Axis in Oral Squamous Cell Carcinoma

Wnt signaling plays a crucial role in the progression of various cancers, including oral squamous cell carcinoma (OSCC). However, the tumor microenvironment (TME) regulating Wnt signaling has not yet been fully elucidated. In this study, we investigated whether cancer-associated fibroblasts (CAFs), the primary components of the TME, activate Wnt signaling and promote tumor progression in OSCC. We conducted a Transwell coculture assay using human OSCC cell lines and normal human dermal fibroblasts (NHDFs). NHDFs stimulated WNT7A expression in several OSCC cell lines, especially HO-1-N-1 and HSC-5. An immunohistochemical study using 122 human OSCC samples indicated that high WNT7A expression in tumor cells was significantly associated with invasion depth and poor prognosis. Moreover, WNT7A expression in OSCC cells was positively correlated with α–smooth muscle actin expression in CAFs. WNT7A knockdown in OSCC cells demonstrated that OSCC cells cocultured with NHDFs significantly promoted tumor cell migration and invasion, which was dependent on WNT7A expression in OSCC cells. We also isolated HSC-5 cells from the coculture and conducted microarray analysis to investigate the factors that promote tumor progression induced by WNT7A. Among the various differentially expressed genes, we identified a downregulated gene encoding CLDN1 and confirmed that WNT7A negatively regulated CLDN1 expression in OSCC cells and CLDN1 knockdown in OSCC cells promoted their migration. Phosphokinase array analysis showed that WNT7A activates protein kinase B (AKT) phosphorylation. Activating AKT signaling using the SC79 agonist induced CLDN1 downregulation in OSCC cells. In the coculture assay, the AKT inhibitor MK2206 significantly recovered CLDN1 expression downregulated by WNT7A, resulting in OSCC cell migration suppression. These results suggest that CAFs stimulate OSCC cells to produce WNT7A, following CLDN1 expression downregulation by activating AKT signaling, promoting cancer cell migration. These findings highlight the importance of molecular therapies targeting the TME in OSCC.

Neutrophils regulate tumor angiogenesis in oral squamous cell carcinoma and the role of Chemerin.

Oral squamous cell carcinoma (OSCC) is the most common malignant tumor of the oral cavity. Tumor angiogenesis plays a crucial role in tumor progression. Studies have established the correlation between neutrophils and tumor angiogenesis in the tumor microenvironment. A previous study found that overexpression of Chemerin- in OSCC increased the infiltration of neutrophils in tumor tissues. This study aims to investigate the mechanisms underlying the regulation of the development and progression of OSCC, which have great significance in enhancing the postoperative survival of patients with OSCC. This study evaluated the accuracy of neutrophil count combined with MVD in predicting patients' survival time and its relationship with clinicopathological parameters and prognosis. Additionally, the study explored the effects of the Chemerin-neutrophil interaction on the angiogenic function of HUVECs. In OSCC, the overexpression of Chemerin promoted the angiogenesis of HUVECs through neutrophils. Moreover, Chemerin upregulated pro-angiogenic factors (e.g., VEGF-A, MMP-9, MMP-2, and S100A9) in neutrophils by activating MEK/ERK signaling pathway. In vivo experiments demonstrated that Chemerin may promote tumor growth by regulating tumor angiogenesis. In conclusion, the results suggest that neutrophil count and MVD serve as poor prognostic factors for patients with OSCC, and their combination is a more effective factor in predicting the survival time of OSCC patients. Neutrophils potentially contribute to angiogenesis through MEK/ERK signaling pathway via Chemerin and participate in the progression and metastasis of OSCC.

MFAP2 promotes the progression of oral squamous cell carcinoma by activating the Wnt/β-catenin signaling pathway through autophagy.

Microfibrillar-associated protein 2 (MFAP2) is a small glycoprotein that is involved in vascular development and metabolic disease. The present study aims to explore the regulatory role of MFAP2 in the development and progression of oral squamous cell carcinoma (OSCC), including the underlying mechanisms. MFAP2 expression and its association with the progression of OSCC are explored using bioinformatics. MFAP2 expression in OSCC tissues is detected by immunohistochemical staining. SCC15 cell migration, invasion, apoptosis, proliferation, and viability are detected by wound healing, Transwell, flow cytometry, colony formation, and cell counting kit-8 assays. An in vivo experiment is used to detect tumor formation. Western blot analysis is used to determine MFAP2's regulatory role in autophagy and the Wnt/β-catenin signaling pathway. MFAP2 is highly expressed in SCC15 cells and OSCC tissues, which correlates positively with the poor prognosis of patients with OSCCs. Functionally, MFAP2 promotes oncogenic autophagy to increase cell invasion, migration, and proliferation but inhibits apoptosis in SCC15 cells and promotes tumor growth in vivo. Mechanistically, MFAP2 upregulates autophagy and Wnt/β-catenin signaling to stimulate OSCC development. Intriguingly, regulation of Wnt/β-catenin signaling dependent on autophagy contributes to the malignant behaviors of SCC15 cells. MFAP2 could serve as a novel biomarker for OSCC and could affect OSCC tumorigenesis and development via autophagic regulation of Wnt/β-catenin signaling.

OTUB1's role in promoting OSCC development by stabilizing RACK1 involves cell proliferation, migration, invasion, and tumor-associated macrophage M1 polarization

Ovarian tumor domain, ubiquitin aldehyde binding 1 (OTUB1), a deubiquitinating enzyme known to regulate the stability of downstream proteins, has been reported to regulate various cancers tumorigenesis, yet its direct effects on oral squamous cell carcinoma (OSCC) progression are unclear. Bioinformatics analysis was performed to screen for genes of interest, and in vitro and in vivo studies were carried out to investigate the function and mechanism of OTUB1 in OSCC. We found that OTUB1 was abnormally elevated in OSCC tissues and positively associated with the pathological stage and tumor stage. Knockdown of OTUB1 impaired the malignance of OSCC cells - suppressed cell proliferation, invasion, migration, and xenografted tumor growth. OTUB1 silencing also drove tumor-associated macrophage M1 polarization but suppressed M2 polarization, and the induction of M1 polarization inhibited the survival of OSCC cells. However, OTUB1 overexpression exerted the opposite effects. Furthermore, the protein network that interacted with the OTUB1 protein was constructed based on the GeneMANIA website. Receptor for activated C kinase 1 (RACK1), a facilitator of OSCC progression, was identified as a potential target of the OTUB1 protein. We revealed that OTUB1 positively regulated RACK1 expression and inhibited RACK1 ubiquitination. Additionally, RACK1 upregulation reversed the effects of OTUB1 knockdown on OSCC progression. Overall, we demonstrated that OTUB1 might regulate OSCC progression by maintaining the stability of the RACK1 protein. These findings highlight the potential roles of the OTUB1/RACK1 axis as a potential therapeutic target in OSCC.

METTL3-Mediated Maturation of miR-99a-5p Promotes Cell Migration and Invasion in Oral Squamous Cell Carcinoma by Targeting ZBTB7A.

METTL3 is an important methyltransferase in N(6)-methyladenosine (m6A) modification. Recently, METTL3 mediates methylation of pri-microRNA (miRNA) to accelerate miRNA maturation, regulating tumor development. This study explored whether METTL3 mediated miR-99a-5p to influence oral squamous cell carcinoma (OSCC) cell metastasis. MiR-99a-5p, ZBTB7A, and MATTL3 expression was measured using quantitative real-time PCR. Biological behaviors were assessed using cell counting kit-8, flow cytometry, Transwell assay, as well as western blot. Luciferase reporter assay evaluated the interaction between miR-99a-5p and ZBTB7A. METTL3-regulated pri-miR-99a-5p processing was determined by RNA binding protein immunoprecipitation (RIP) and methylated RNA immunoprecipitation (MeRIP) assays. The consequences clarified that miR-99a-5p was upregulated in OSCC cells. Downregulation of miR-99a-5p suppressed cellular viability, migration, invasion, and epithelial-mesenchymal transition (EMT), and induced apoptosis. ZBTB7A acted as a miR-99a-5p target and reversed the effects on cellular behaviors induced by miR-99a-5p inhibitor. m6A content and METTL3 expression were increased in OSCC cells. METTL3 promoted the m6A modification of pri-miR-99a-5p and thereby facilitated miR-99a-5p processing. Moreover, knockdown of METTL3 inhibited OSCC metastasis by downregulating miR-99a-5p. Taken together, METTL3 promoted miR-99a-5p maturation in an m6A-dependent manner, which further targets ZBTB7A to accelerate the progression of OSCC. These findings suggest potential targets for OSCC therapy.

Microvascular density analysis and histological parameters of oral cancer progression.

This study aimed to investigate the role of blood and lymphatic microvascular density in the progression of oral squamous cell carcinoma (OSCC). The sample was composed of 54 cases of OSCC. The immunoexpression to anti-alpha-smooth muscle actin (α-SMA) and to anti-endoglin (CD105) was used to determine the microvessel density (MVD); anti-podoplanin (D2-40) was used to assess the lymphatic vessel density (LVD); vascular endothelial growth factor (VEGF) was evaluated in malignant cells. The histological differentiation, the worst pattern of invasion (WPOI), tumour thickness and tumour budding (TB) intensity were assessed using haematoxylin-eosin and anti-pan-cytokeratin (AE1/AE3). Patients' age and sex, TNM classification and follow-up time were collected from the medical records. MVD markers presented a similar pattern of expression in blood vessels. However, only α-SMA + MVD was significantly higher among women and in tumours ≤4 cm. LVD was lower in tumours with lymph node metastasis. Regarding the histological parameters, high TB intensity was associated with histological differentiation, advanced clinical stage, greater tumour thickness and reduced disease-free survival. No difference was found in VEGF. The decrease in OSCC LVD could be related to pathological node involvement, whereas high TB intensity could indicate OSCC progression and worse patient outcomes.

Targeting Circ-OCAC suppress oral squamous cell carcinoma progression.

Circular RNAs (circRNAs), with their multilevel and versatile regulation, have emerged as promising targets for treating complex and heterogeneous malignancies such as oral squamous cell carcinoma (OSCC). It is crucial to explore the function of key circRNAs and elucidate the underlying mechanisms to establish an effective in vivo delivery system to better utilize circRNAs as cancer treatment strategies. circRNA (circ-OCAC) was identified as significantly downregulated in tumor samples compared to paracancerous tissues by RNA-seq analysis of eight pairs of OSCC tissues. Functional experiments of circ-OCAC were performed both in vitro and in vivo. The interactions between circ-OCAC and miR-411-5p were clarified by RNA pull down and RNA immunoprecipitation (RIP) assays. We observed that circ-OCAC inhibits OSCC growth and metastasis by blocking the PI3K/Akt signaling pathway. To translate this observation in vivo, a pH-responsive nanoparticle (pNP) was developed to target circ-OCAC. Our results confirmed the advantages of the pNP-circ-OCAC system: high tumor enrichment capacity and good biosafety, which resulted in a significantly enhanced antitumor effect. This study demonstrated that targeting circ-OCAC serves as a promising potential therapeutic strategy for OSCC.

Expression levels of microRNA-7110 in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is a leading cause of cancer-related deaths worldwide, and it is responsible for more than 95% of head and neck cancers. Despite advancements in research and treatment, patient's survival has not significantly increased in recent years. On the other hand, microRNAs (miRNAs) are a major class of small non-coding RNAs that regulate gene expression of the target mRNAs. Thus, understanding the mechanisms behind OSCC formation and progression may lead to the identification of potential diagnostic biomarkers and therapeutic molecules for the treatment of OSCC. The aim of the current study was to analyze expression levels of miR-7110 in OSCC tissues and adjacent normal tissues as it could provide insights into its potential role in OSCC development or progression as a valuable biomarker. A total of 20 OSCC and adjacent normal tissues were collected from the Department of Oral and Maxillofacial Surgery, Saveetha Dental College and Hospitals (Chennai, India). The tissues were processed for hematoxylin and eosin staining and expression studies. The data were shown as mean±standard deviation and P<0.05 was considered statistically significant. Our histopathological observations revealed an invasive malignant epithelial neoplasm with malignant epithelial cells exhibiting features of severe epithelial dysplasia invading the connective tissue stroma as islands, strands and cords with varying degrees of differentiation. Our results have also revealed that the expression levels of miR-7110 were found to be significantly higher in OSCC samples when compared to the normal tissue. We can preliminarily conclude that based on the increased expression of miR-7110 in OSCC tissue samples, they can be used as an early diagnostic or prognostic biomarker and/or a therapeutic target for the treatment of OSCC even though more focused research in that direction is needed.

Prognostic model based on telomere-related genes predicts the risk of oral squamous cell carcinoma

BackgroundThis study investigated a potential prognostic model based on telomere-related genes (TRGs) for the clinical prediction of oral squamous cell carcinoma (OSCC).MethodsGene expression data and associated clinical phenotypes were obtained from online databases. Differentially expressed (DE)-TRGs were identified between OSCC and normal samples, followed by protein-protein interaction and enrichment analyses. Subsequently, the prognostic genes explored based on the DE-TRGs and survival data were applied in the establishment of the current prognostic model, and an integrated analysis was performed between high- and low-risk groups using a prognostic model. The expression of certain prognostic genes identified in the present study was validated using qPCR analysis and/or western blot in OSCC cell lines and clinical samples.Results169 DE-TRGs were identified between the OSCC samples and controls. DE-TRGs are mainly involved in functions such as hypoxia response and pathways such as the cell cycle. Eight TRGs (CCNB1, PDK4, PLOD2, RACGAP1, MET, PLK1, KPNA2, and CCNA2) associated with OSCC survival and prognosis were used to construct a prognostic model. qPCR analysis and western blot showed that most of the eight prognostic genes were consistent with the current bioinformatics results. Analysis of the high- and low-risk groups for OSCC determined by the prognostic model showed that the current prognostic model was reliable.ConclusionsA novel prognostic model for OSCC was constructed by TRGs. PLOD2 and APLK1 may participate in the progression of OSCC via responses to hypoxia and cell cycle pathways, respectively. TRGs, including KPNA2 and CCNA2, may serve as novel prognostic biomarkers for OSCC.

Identification and validation of genes associated with copper death in oral squamous cell carcinoma based on machine learning and weighted gene co-expression network analysis

ObjectiveTo identify copper-induced death-associated hub genes in oral squamous cell carcinoma (OSCC) and understand their functional and biological significance using machine learning and Weighted Gene Co-expression Network Analysis (WGCNA). MethodsOSCC transcriptomic data from GEO and TCGA databases were subjected to data integration, batch effect removal, background correction, and quantile normalization to select cuproptosis-associated genes using Spearman's correlation analysis. The 'limma' R package was used to filter differentially expressed genes (DEGs). Core module genes selected using gene co-expression network analysis with R package 'WGCNA' were screened using Support Vector Machine (SVM), LASSO regression, and Random Forest (RF) machine learning algorithms and validated using TCGA database samples. Core gene expression variations between OSCC and adjacent normal tissues were validated using immunohistochemistry. Immune infiltration analysis using package 'CIBERSORT' correlated hub genes with immune cells. ResultsFrom 19 cuproptosis-related genes (identified from literature), 2382 cuproptosis-related mRNA were obtained through Spearman's correlation analysis; 112 DEGs using 'limma' R package and 32 hub genes using WGCNA were obtained. Hub genes TMPRSS11B, SERPINH1, and CDH3 were identified using machine learning algorithms. TCGA validation showed that TMPRSS11B significantly underexpressed (P < 0.001) but SERPINH1 and CDH3 significantly overexpressed (P < 0.001) in tumor samples. The AUC for TMPRSS11B, SERPINH1, and CDH3 in ROC curve analysis were 78.1%, 95.6%, and 87.5%, respectively. ConclusionTMPRSS11B, SERPINH1, and CDH3 may be pivotal for OSCC development and progression and potential targets for new therapeutic and predictive strategies. However, their specific functions and mechanisms underlying OSCC remain to be elucidated.

Long non-coding RNA SLC7A11 antisense RNA1 promotes oral squamous cell carcinoma progression by regulating ubiquitination of K-homology type splicing regulatory protein

ObjectivesThis article aims to elucidate the role of Long non-coding RNA SLC7A11 antisense RNA1 (SLC7A11-AS1) in oral squamous cell carcinoma, which are expected to be useful for the oral squamous cell carcinoma diagnosis and treatment. DesignSLC7A11-AS1 expression was detected by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) in oral squamous cell carcinoma cell lines. Cellular localization of SLC7A11-AS1C was detected by fluorescence in situ hybridization (FISH) assays and subcellular fractionation assay. Biological functions of SLC7A11-AS1 were explored by 3-(4,5-dimethyl-2-thiazolyl)− 2,5-diphenyl-2-H-tetrazolium bromide (MTT), 5-ethynyl-2′-deoxyuridine (EdU), wounding healing, and transwell invasion assays in vitro, as well as mice xenograft experiments and metastasis assays in vivo. RNA pull-down, RNA immunoprecipitation, co-immunoprecipitation, ubiquitination assays, and rescue experiments were performed to determine the molecular mechanism of SLC7A11-AS1 in oral squamous cell carcinoma. ResultsSLC7A11-AS1 is overexpressed in oral cancer tissues and cell lines. Functionally, knockdown of SLC7A11-AS1 reduced the proliferation, migration, and invasion of oral squamous cell carcinoma cells in vitro and inhibited tumor growth as well as metastasis in vivo. Mechanistically, SLC7A11-AS1 impeded the interaction between K-homology type splicing regulatory protein (KHSRP) and kelch-like 12 (KLHL12), maintaining the stability of KHSRP by restraining KHSRP degradation through the ubiquitination-proteasome pathway. Furthermore, KHSRP overexpression recovered the malignant behaviors inhibited by SLC7A11-AS1 knockdown in oral cancer cells. ConclusionSLC7A11-AS1 promoted oral squamous cell carcinoma development by interacting with KHSRP and maintaining KHSRP stability by preventing its degradation via the ubiquitination-proteasome pathway. Thus, SLC7A11-AS1 is a potential therapeutic target for oral cancer.

LncRNA BANCR promotes oral squamous cell carcinoma progression via regulating Rab1A signaling.

Long non-coding RNA BRAF-activated non-protein coding RNA plays bidirectional roles in human cancers. However, function and molecular mechanism of BRAF-activated non-protein coding RNA in oral squamous cell carcinoma still need to clarify further. Long non-coding RNA microarray assay, in situ hybridization staining, clinicopathological data analysis were performed to investigate expression pattern of BRAF-activated non-protein coding RNA in oral squamous cell carcinoma tissue samples. Constructing ectopically expressed BRAF-activated non-protein coding RNA in oral squamous cell carcinoma cells via plasmids or siRNAs, then changeable abilities of proliferation and motility of these cells were observed in vitro and in vivo. RNA-protein pulldown, RNA immunoprecipitation, and bioinformatics analyses were performed to explore potential pathways involved in BRAF-activated non-protein coding RNA-based regulation of malignant progression in oral squamous cell carcinoma. BRAF-activated non-protein coding RNA was identified upregulated in oral squamous cell carcinoma tissue and correlated with nodal metastasis and clinical severity of patients. Overexpressed BRAF-activated non-protein coding RNA increased percentage of 5-ethynyl-2'-deoxyuridine-positive cells, viability, migration, and invasion rates of oral squamous cell carcinoma cells, while silenced BRAF-activated non-protein coding RNA could observe weakened effects in vitro. Xenograft tumor formed by BRAF-activated non-protein coding RNA-overexpressed cells had bigger volume, faster growth rates, higher weight, and more Ki67+ cells. Pulmonary metastasis induced by BRAF-activated non-protein coding RNA-silenced cells had fewer colony nodes, Ki67+ cells, and CD31+ blood vessels. Furthermore, BRAF-activated non-protein coding RNA was mainly localized in nucleus of oral squamous cell carcinoma cells and bound Ras-associated binding 1A. Silencing Ras-associated binding 1A could damage mobile ability and phosphorylation levels of nuclear factor-κBin oral squamous cell carcinoma cells induced by overexpressing BRAF-activated non-protein coding RNA. Opposite trend was also observed. Acting as a promoter in oral squamous cell carcinoma metastasis, BRAF-activated non-protein coding RNA promotes oral squamous cell carcinoma cells proliferation and motility by regulating the BRAF-activated non-protein coding RNA/Ras-associated binding 1A complex, which activates nuclear factor-κBsignaling pathway.

Yin Yang 1-Induced Long Noncoding RNA DUXAP9 Drives the Progression of Oral Squamous Cell Carcinoma by Blocking CDK1-Mediated EZH2 Degradation.

LncRNAs play a critical role in oral squamous cell carcinoma (OSCC) progression. However, the function and detailed molecular mechanism of most lncRNAs in OSCC are not fully understood. Here, a novel nuclear-localized lncRNA, DUXAP9 (DUXAP9), that is highly expressed in OSCC is identified. A high level of DUXAP9 is positively associated with lymph node metastasis, poor pathological differentiation, advanced clinical stage, worse overall survival, and worse disease-specific survival in OSCC patients. Overexpression of DUXAP9 significantly promotes OSCC cell proliferation, migration, invasion, and xenograft tumor growth and metastasis, and upregulates N-cadherin, Vimentin, Ki67, PCNA, and EZH2 expression and downregulates E-cadherin in vitro and in vivo, whereas knockdown of DUXAP9 remarkably suppresses OSCC cell proliferation, migration, invasion, and xenograft tumor growth in vitro and in vivo in an EZH2-dependent manner. Yin Yang 1 (YY1) is found to activate the transcriptional expression of DUXAP9 in OSCC. Furthermore, DUXAP9 physically interacts with EZH2 and inhibits EZH2 degradation via the suppression of EZH2 phosphorylation, thereby blocking EZH2 translocation from the nucleus to the cytoplasm. Thus, DUXAP9 can serve as a promising target for OSCC therapy.