human oscc cell lines cal27  (ATCC)

Journal: International Dental Journal

Article Title: Waterpipe Smoke-Induced N6-Methyladenosine Modification Promotes the Progression of Oral Squamous Cell Carcinoma

doi: 10.1016/j.identj.2025.109295

Figure Lengend Snippet: WPSC induces METTL3 and its target oncogene HMGA2 expression in OSCC cells. WPSC induces METTL3 mRNA expression (A) and its target HMGA2 mRNA expression (B) in OSCC cells (CAL27) ( P < .001). WPSC induces METTL3 and its target HMGA2 protein expression in OSCC cells (C). METTL3 expression positively correlated with its target HMGA2 expression (D). The SRAMP database showed that HMGA2 mRNA was enriched with m6A methylation. (E). Statistical significance is denoted as *** P < .001.

Article Snippet: The human OSCC cell line CAL27 was obtained from the American Type Culture Collection (ATCC, USA) and cultured in DMEM supplemented with 10% FBS and antibiotics.

Techniques: Expressing, Methylation

Journal: International Dental Journal

Article Title: Waterpipe Smoke-Induced N6-Methyladenosine Modification Promotes the Progression of Oral Squamous Cell Carcinoma

doi: 10.1016/j.identj.2025.109295

Figure Lengend Snippet: METTL3 expression analysis. METTL3 mRNA expression increased in primary and metastatic OSCC patients (A). METTL3 protein expression increased in primary and metastatic OSCC patients (B). TCGA-OSCC dataset showed that METLL3 mRNA increased in the oral cavity (C), tongue (D), and metastatic OSCC patients (E). Statistical significance is denoted as * P < .05, *** P < .001.

Article Snippet: The human OSCC cell line CAL27 was obtained from the American Type Culture Collection (ATCC, USA) and cultured in DMEM supplemented with 10% FBS and antibiotics.

Techniques: Expressing

Journal: BMC Oral Health

Article Title: Expression profile of GSDMB in oral squamous cell carcinoma and its impact on tumor immune microenvironment and prognosis

doi: 10.1186/s12903-026-07781-1

Figure Lengend Snippet: Expression landscape of GSDMB across different groups (330 OSCC tissues, 32 normal tissues). A - C Differential expression of GSDMB among Grade categories, age groups, and N stages. D Boxplot showing the expression difference of GSDMB between OSCC cases and controls. E Receiver Operating Characteristic (ROC) curve for GSDMB. F Forest plot from univariate Cox regression analysis based on GSDMB expression levels and prognostic outcomes across various cancers. G Boxplot illustrating differences in GSDMB expression between control and disease groups across pan-cancer samples. “wilcox.test” is used to calculate the significance p-value (* p<0.05, ** p<0.01, *** p<0.001)

Article Snippet: The human OSCC cell lines SCC4 and SCC25 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Quantitative Proteomics, Control

Journal: BMC Oral Health

Article Title: Expression profile of GSDMB in oral squamous cell carcinoma and its impact on tumor immune microenvironment and prognosis

doi: 10.1186/s12903-026-07781-1

Figure Lengend Snippet: Landscape of Tumor Mutational Burden (TMB) in high and low GSDMB expression groups (318 OSCC tissues). A - B Waterfall plots depicting mutations in high and low GSDMB expression groups. C Boxplot comparing TMB differences between the two groups. D Scatter plot of the correlation between TMB and GSDMB expression levels. E Kaplan-Meier survival curves for high and low TMB groups. F KM survival curves for four sample groups classified by combined TMB and GSDMB expression levels

Article Snippet: The human OSCC cell lines SCC4 and SCC25 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing

Journal: BMC Oral Health

Article Title: Expression profile of GSDMB in oral squamous cell carcinoma and its impact on tumor immune microenvironment and prognosis

doi: 10.1186/s12903-026-07781-1

Figure Lengend Snippet: Analysis of differences between the two groups and GSEA analysis results (330 OSCC tissues). A Volcano plot displaying DEGs. B Heatmap of DEGs illustrating expression levels across samples. C Circle plot combined with a network view of GO/KEGG pathway enrichment analysis. D Bar plot summarizing the results of GO/KEGG enrichment analysis. E - L Gene set enrichment analysis (GSEA) results for key KEGG pathways. Specific pathways include ECM-receptor interaction ( E ), focal adhesion ( F ), MAPK signaling ( G ), natural killer cell-mediated cytotoxicity ( H ), primary immunodeficiency ( I ), T-cell receptor signaling ( J ), TGF-beta signaling ( K ), and Toll-like receptor signaling ( L ). Key statistics (NES: Normalized Enrichment Score; p-value; FDR: False Discovery Rate) are provided in the plots

Article Snippet: The human OSCC cell lines SCC4 and SCC25 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing

Journal: BMC Oral Health

Article Title: Expression profile of GSDMB in oral squamous cell carcinoma and its impact on tumor immune microenvironment and prognosis

doi: 10.1186/s12903-026-07781-1

Figure Lengend Snippet: Basic analysis results of scRNA-seq data. A Violin plots of six OSCC samples' scRNA-seq data post-merging and quality control, indicating the number of nonzero-expressed genes per cell, total UMI counts, and mitochondrial gene percentage. B Two-dimensional distribution of six samples with batch effects. C Distribution of different samples after removing batch effects using the "Harmony" package in R. D Heatmap predicting cell cluster identities using the "singleR" software package. E Two-dimensional distribution of different cell types. F Expression of GSDMB in different cell types. G Distribution of six OSCC samples across different cell types

Article Snippet: The human OSCC cell lines SCC4 and SCC25 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Control, Software, Expressing

Journal: BMC Oral Health

Article Title: Expression profile of GSDMB in oral squamous cell carcinoma and its impact on tumor immune microenvironment and prognosis

doi: 10.1186/s12903-026-07781-1

Figure Lengend Snippet: Classification of high and low expression cell clusters and GSDMB expression in stRNA-seq data. A All cells were divided into high and low expression clusters based on the median value of GSDMB. B Proportions of different cell types within high and low expression clusters. C Results of GSEA analysis for the two clusters. D Clustering results for four slices based on OSCC stRNA-seq data. E Cell type annotation results for four slices from OSCC stRNA-seq data. F Expression of GSDMB in different cell types

Article Snippet: The human OSCC cell lines SCC4 and SCC25 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing

Journal: BMC Oral Health

Article Title: Expression profile of GSDMB in oral squamous cell carcinoma and its impact on tumor immune microenvironment and prognosis

doi: 10.1186/s12903-026-07781-1

Figure Lengend Snippet: GSDMB silencing attenuates the malignant properties and modulates 5-FU induced pyroptosis related responses in SCC25 cells. A IHC analysis of GSDMB expression in OSCC tissues and normal oral mucosa (n = 40). B – C Western blot analysis of GSDMB protein levels in HOK and OSCC cell lines and in SCC25 cells following siRNA transfection. D Cell viability assessed by the CCK-8 assay. E Representative images and quantification of colony formation. F Wound healing assay evaluating cell migration. G Transwell invasion assay. H Dose–response curves and corresponding IC50 values for 5-fluorouracil (5-FU). I Lactate dehydrogenase (LDH) release assay assessing 5-FU induced lytic cell death. J – K ELISA quantification of IL-1β and IL-18 levels in culture supernatants after 5-FU treatment. (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The human OSCC cell lines SCC4 and SCC25 were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Western Blot, Transfection, CCK-8 Assay, Wound Healing Assay, Migration, Transwell Invasion Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: GINS2 promotes oral squamous cell carcinoma progression and immune evasion by recruiting PD-L1 + neutrophils and modulating the PTP4A1/PKM2 axis

doi: 10.3389/fimmu.2025.1637296

Figure Lengend Snippet: GINS2 is upregulated in OSCC and correlates with adverse clinicopathological features. (a) Volcano plot showing differentially expressed genes between HNSC tumor tissues and normal tissues from TCGA database. (b) STRING PPI network of prognostic DEGs (|log2FC| > 2; FDR < 0.05; survival-associated). Genes not present in the seed (e.g., PTP4A1, PKM) are not displayed. The panel is intended to position GINS2 within replication/cell-cycle interactors. (c) Pan-cancer analysis from TCGA showing GINS2 mRNA expression (log2 TPM) across cancer types (red) versus normal tissues (blue). (d) Representative IHC images (left) and quantification (right) of GINS2 expression in OSCC versus controls. Scale bar = 50 µm. (e) Relative GINS2 mRNA expression stratified by clinical stage (qPCR; OSCC patient samples). (f) Relative GINS2 mRNA expression stratified by pathological grade (qPCR; OSCC patient samples). (g) Western blot (left) and quantification (right) demonstrating GINS2 protein expression in multiple OSCC cell lines (Cal27, HN6, SCC4, SCC25) compared to normal human oral keratinocytes (HOK). β-actin served as the loading control. Data in plots are presented as mean ± SD or box plots showing median and interquartile range. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Human OSCC cell lines (Cal27, HN6, SCC4, SCC25) and normal human oral keratinocytes (HOK) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: GINS2 promotes oral squamous cell carcinoma progression and immune evasion by recruiting PD-L1 + neutrophils and modulating the PTP4A1/PKM2 axis

doi: 10.3389/fimmu.2025.1637296

Figure Lengend Snippet: GINS2 interacts with PTP4A1 and regulates its expression, and PTP4A1 interacts with PKM2 in OSCC cells. (a) Western blot analysis showing baseline protein expression of PTP4A1 in normal human oral keratinocytes (HOK) and OSCC cell lines HN6 and SCC25. β-actin served as the loading control. Bar graph shows quantification relative to HOK. (b, c) Western blot analysis (top) and quantification (bottom) demonstrating the effect of GINS2 knockdown (sh-GINS2) or overexpression (OE-GINS2) on PTP4A1 protein levels in HN6 (b) and SCC25 (c) cells, compared to respective controls (sh-NC, OE-NC). β-actin served as the loading control. (d) Co-immunoprecipitation (Co-IP) assays. Left panels: Interaction between endogenous GINS2 and PTP4A1 in HN6 cells. IP with anti-GINS2 pulls down PTP4A1, and IP with anti-PTP4A1 pulls down GINS2. Right panels: Interaction between endogenous PTP4A1 and PKM2 in HN6 and SCC25 cells. IP with anti-PTP4A1 pulls down PKM2. Control IgG IPs and input lysates are shown. (e) Immunofluorescence staining showing co-localization of PTP4A1 (green) and PKM2 (red) in HN6 and SCC25 cells. Nuclei are stained with DAPI (blue). Scale bar = 20 µm. Data in bar graphs are presented as mean ± SD. ***P < 0.001.

Article Snippet: Human OSCC cell lines (Cal27, HN6, SCC4, SCC25) and normal human oral keratinocytes (HOK) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Western Blot, Control, Knockdown, Over Expression, Immunoprecipitation, Co-Immunoprecipitation Assay, Immunofluorescence, Staining

Journal: Frontiers in Immunology

Article Title: GINS2 promotes oral squamous cell carcinoma progression and immune evasion by recruiting PD-L1 + neutrophils and modulating the PTP4A1/PKM2 axis

doi: 10.3389/fimmu.2025.1637296

Figure Lengend Snippet: GINS2 promotes OSCC proliferation, migration, invasion, and tumorigenesis. (a) qPCR confirming efficient knockdown of GINS2 mRNA in HN6 and SCC25 cells after sh-GINS2 transfection compared to sh-NC. β-actin was used for normalization. (b) CCK-8 assay over 96 hours showing reduced viability of HN6 and SCC25 cells upon GINS2 knockdown. (c) Colony formation assay images (top) and quantification (bottom) demonstrating fewer and smaller colonies after GINS2 silencing. (d) Wound-healing assay images (0 h, 12 h, 24 h, and 48 h) and quantification showing impaired migration. Scale bar = 200 µm. (e) Transwell Matrigel invasion assay images (left) and quantification (right) showing reduced invasion after GINS2 silencing. Invaded cells stained with crystal violet. Scale bar = 100 µm. (f–h) Effect of GINS2 knockdown on OSCC tumor growth in vivo . Representative xenograft images (f) , tumor growth curves (g) , and final tumor weight (h) showing suppressed tumorigenesis in sh-GINS2 vs sh-NC HN6 cells. Data are presented as mean ± SD. ***P < 0.001. n = 5 mice per group.

Article Snippet: Human OSCC cell lines (Cal27, HN6, SCC4, SCC25) and normal human oral keratinocytes (HOK) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Migration, Knockdown, Transfection, CCK-8 Assay, Colony Assay, Wound Healing Assay, Invasion Assay, Staining, In Vivo

Journal: Frontiers in Immunology

Article Title: GINS2 promotes oral squamous cell carcinoma progression and immune evasion by recruiting PD-L1 + neutrophils and modulating the PTP4A1/PKM2 axis

doi: 10.3389/fimmu.2025.1637296

Figure Lengend Snippet: GINS2 expression correlates with neutrophil infiltration, GINS2 promotes TAN accumulation, and neutrophil depletion impairs GINS2-driven tumor growth. (a) Correlation analyses using TIMER2.0 database in the HNSC cohort. Left: Scatter plot showing correlation between GINS2 expression level (log2 TPM) and estimated tumor purity. Right: Scatter plot showing correlation between GINS2 expression level and neutrophil infiltration level estimated by the TIMER algorithm. Spearman’s rho and P-value are indicated. (b) Left: Representative immunohistochemistry (IHC) staining for Myeloperoxidase (MPO) in control oral tissue and OSCC tissue, showing increased neutrophil infiltration in OSCC. Right: Quantification of MPO expression (% Area) from IHC images (n=5 per group). Scale bar = 50 µm. (c) Left: Representative immunofluorescence images showing staining for MPO (red) and citrullinated histone H3 (CitH3, green) in OSCC tissue sections. Nuclei are stained with DAPI (blue). Merged images suggest potential NET formation. Right: corresponding quantification. Scale bar = 50 µm. (d) Flow cytometry analysis quantifying the percentage of CD11b+Ly6G+ neutrophils within subcutaneous xenograft tumors derived from HN6 cells transfected with sh-NC, sh-GINS2, OE-NC, or OE-GINS2. Gating strategy: live (viability dye - ) → CD45 + leukocytes → CD11b + Ly6G + neutrophils. Readout: %CD11b + Ly6G + of CD45 + cells within the tumor digest. Representative flow plots (left) and quantification (right) are shown. (e) Effect of neutrophil depletion on GINS2-driven tumor growth in vivo . Representative images of subcutaneous xenograft tumors derived from OE-GINS2 HN6 cells in nude mice treated with control IgG or anti-Ly6G antibody (left), and quantification of tumor volume (middle) over time and final tumor weight (right). Data are presented as mean ± SD. **P < 0.01, ***P < 0.001.

Article Snippet: Human OSCC cell lines (Cal27, HN6, SCC4, SCC25) and normal human oral keratinocytes (HOK) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Immunohistochemistry, Control, Immunofluorescence, Staining, Flow Cytometry, Derivative Assay, Transfection, In Vivo

Journal: Frontiers in Immunology

Article Title: GINS2 promotes oral squamous cell carcinoma progression and immune evasion by recruiting PD-L1 + neutrophils and modulating the PTP4A1/PKM2 axis

doi: 10.3389/fimmu.2025.1637296

Figure Lengend Snippet: GINS2-associated neutrophils express PD-L1, mediating immunosuppression, and targeting GINS2 synergizes with anti-PD-L1 therapy in vivo . (a) Left: Representative flow cytometry histogram showing higher PD-L1 surface expression (MFI) on neutrophils from OSCC environment compared to control. (a) Right: Dot plot showing a significant positive correlation between GINS2 relative expression (in matched tumors) and PD-L1 MFI on associated neutrophils (n = 15 matched OSCC cases, Spearman correlation). Gating strategy: live (viability dye - ) → CD66b + neutrophils → PD-L1 (PE-conjugated) histogram/overlay. Readouts: PD-L1 MFI and %PD-L1 + among CD66b + cells (FMO-anchored). (b) EdU incorporation assay assessing proliferation of CD8+ T cells co-cultured with neutrophils +/- anti-PD-L1 antibody or control IgG. Representative images (upper) and quantification (lower) show increased T cell proliferation upon PD-L1 blockade. Scale bar = 50 µm. (c) Flow cytometry analysis assessing apoptosis (Annexin V-FITC/PI staining) of CD8+ T cells after co-culture and treatment with control IgG or anti-PD-L1 antibody. Upper: Representative flow plots. Lower: Quantification of the percentage of apoptotic (Annexin V+) CD8+ T cells. Gating strategy: lymphocytes (FSC-A vs SSC-A) → singlets (FSC-H/FSC-A) → live (viability dye - ) → CD3 + CD8 + T cells → Annexin V-FITC vs PI. Readouts: %Annexin V + (early + late apoptosis) among CD3 + CD8 + cells; PI used to distinguish early (Annexin V + /PI - ) vs late (Annexin V + /PI + ) apoptosis when reported. (d) Representative immunofluorescence images showing expression of PD-1 (red) and TIM-3 (green) on cells (likely T cells) within the tumor microenvironment. Nuclei are stained with DAPI (blue). Scale bar = 50 µm. (e) In vivo therapeutic efficacy study using NOD/SCID mice reconstituted with human T cells and neutrophils, injected with sh-GINS2 HN6 cells, and treated with PBS, control IgG, or anti-PD-L1 antibody. Representative images of excised tumors (upper), and quantification of tumor volume over time (middle) and final tumor weight (lower) across different groups: PBS, T cell only, T cell+neutrophils, T cell+neutrophils+sh-GINS2, T cell+neutrophils+sh-GINS2+anti-PD-L1. (f) Representative IHC staining for Ki67 in xenograft tumors from the different treatment groups in the in vivo study described in (d) . Scale bar = 50 µm. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Human OSCC cell lines (Cal27, HN6, SCC4, SCC25) and normal human oral keratinocytes (HOK) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vivo, Flow Cytometry, Expressing, Control, Cell Culture, Staining, Co-Culture Assay, Immunofluorescence, Drug discovery, Injection, Immunohistochemistry