human oscc cell lines scc4  (ATCC)

Journal: Frontiers in Oncology

Article Title: MUC21 is downregulated in oral squamous cell carcinoma and associated with poor prognosis

doi: 10.3389/fonc.2026.1767261

Figure Lengend Snippet: MUC21 Expression by analyzing high-throughput dataset including Affymetrix GeneChip ® Human Transcriptome Array 2.0(HTA), GSE34105 from GEO and TCGA RNAseq data. (A-F) , the heatmaps of hierarchical clustering and volcano plots of differential gene expressions revealed by HTA based on 10 paired OSCC and adjacent normal oral tissue; GSE34105 based on 62 OSCC and 16 normal oral tissue; TCGA base on 266 OSCC and 19 normal oral tissue. Genes with a fold change >2 and adj-P-value of <0.05 were highlighted. (G, H) the up and down regulated genes in the three datasets were intersected and it was showed that 73 was up and 102 were down regulated in the intersection which comprised only MUC15 and MUC21 from Mucin family. (I) relative expression level (the median of the three datasets) comparison showed that MUC21 was more down regulated than MUC15, the chart was created from Excel (Microsoft ® for Excel). OSCC, oral squamous cell carcinoma.

Article Snippet: Human OSCC cell lines Cal27 and HN6 were acquired from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, High Throughput Screening Assay, RNA sequencing, Comparison

Journal: Frontiers in Oncology

Article Title: MUC21 is downregulated in oral squamous cell carcinoma and associated with poor prognosis

doi: 10.3389/fonc.2026.1767261

Figure Lengend Snippet: Quantitative qRT-PCR and immunohistochemistry analysis of MUC21, KRT4, KRT13, and CRNN in OSCC and para-OSCC. (A-D) showed that MUC21, KRT4, KRT13 and CRNN were down regulated in OSCC than para-OSCC (P< 0.0001), the expression levels were normalized against GAPDH. (E) Spearman correlation analysis showed that MUC21was related with KRT4, KRT13 and CRNN at mRNA level. (F-I) showed that MUC21 and KRT4 were down regulated in OSCC than para-OSCC (P<0.0001); KRT13 was down regulated in OSCC than para-OSCC (P <0.05); and CRNN was also down regulated in OSCC than para-OSCC (P<0.001). (J) Spearman correlation analysis showed that MUC21 was related with KRT4, KRT13 and CRNN at protein level too.

Article Snippet: Human OSCC cell lines Cal27 and HN6 were acquired from the American Type Culture Collection (ATCC, USA).

Techniques: Quantitative RT-PCR, Immunohistochemistry, Expressing

Journal: Frontiers in Oncology

Article Title: MUC21 is downregulated in oral squamous cell carcinoma and associated with poor prognosis

doi: 10.3389/fonc.2026.1767261

Figure Lengend Snippet: MUC21 expression analysis in OSCC and para-OSCC via immunohistochemistry (IHC) and its relation with critical clinical characters. (A) a whole block of OSCC and para-OSCC tissue analyzed by IHC showed that MUC21 was expressed in para-OSCC epithelium and lost in OSCC. (B) MUC21 expression between OSCC and para-OSCC in 102 paired patient samples was quantified by mean optical density (MOD) values. MUC21 decreased significantly in OSCC (P < 0.0001). Box plots represent the median, 25th, and 75th percentiles of the data. (C–H) displayed matched para-OSCC and OSCC. (D, F, H) represented well, moderate, and poor differentiation of OSCC, respectively. Unannotated Scale bar = 100 μm. The Scale bar of the block tissue was 1mm. (I) decreased MUC21 expression level was related with cervical lymphatic metastasis and OSCC differentiation. “pN0” means no lymphatic metastasis, “pN1-PN3” referred to different degrees of lymphatic metastasis; “well +moderate” and “poor” referred to different degrees of differentiation.

Article Snippet: Human OSCC cell lines Cal27 and HN6 were acquired from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Immunohistochemistry, Blocking Assay

Journal: Frontiers in Oncology

Article Title: MUC21 is downregulated in oral squamous cell carcinoma and associated with poor prognosis

doi: 10.3389/fonc.2026.1767261

Figure Lengend Snippet: Kaplan–Meier survival analyses for postoperative OSCC patients based on MUC21 expression. (A, B) Overall Survival (OS) and disease-free survival (DFS)with low MUC21 expression were significantly shorter than those with high MUC21 expression (P = 0.012 for OS, P<0.0001 for DFS). (C, D) , Forest map: The univariate analysis of OS and DFS in OSCC patients. (E, F) , Forest map: The multivariate analysis of OS and DFS in OSCC patients.

Article Snippet: Human OSCC cell lines Cal27 and HN6 were acquired from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: MUC21 is downregulated in oral squamous cell carcinoma and associated with poor prognosis

doi: 10.3389/fonc.2026.1767261

Figure Lengend Snippet: In vitro cell lines experiment post overexpression and knockdown of MUC21. (A) MUC21 was significantly overexpressed and knocked down in CAL27 and HN6. (B) CCK-8 assay on CAL27 and HN6 post MUC21 manipulation. (C) Transwell assay without Matrigel coated filter of CAL27 and HN6 post MUC21 manipulation. (D) Transwell assay with Matrigel coated filter of CAL27 and HN6 post MUC21 manipulation. (E) wound-healing assay conducted at 24 hours post MUC21 manipulation in CAL27 and HN6 cells. NC: negative control, SH: MUC21 knockdown, OE: MUC21 overexpression. Statistical significance is denoted by *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: Human OSCC cell lines Cal27 and HN6 were acquired from the American Type Culture Collection (ATCC, USA).

Techniques: In Vitro, Over Expression, Knockdown, CCK-8 Assay, Transwell Assay, Wound Healing Assay, Negative Control

Journal: Cell Death & Disease

Article Title: Histone lactylation-driven feedback loop modulates pyrimidine metabolism to promote oral carcinogenesis

doi: 10.1038/s41419-026-08580-w

Figure Lengend Snippet: A UMAP plot of the clustering results for eight major cell types from OLK and OSCC tissues. B Marker genes of each cluster. C UMAP plot showing the cell distribution across all 14 samples. D Reactome pathway analysis of DEGs with gradually increased expression between epithelial cells from OLK and primary OSCC. E Lactate content in normal ( n = 30), OLK ( n = 30), and OSCC ( n = 30) tissues. F Lactate content in HOK, DOK, and OSCC cells (HSC3, HN4, HN6, and CAL27). The representative images (×200) ( G ) and quantification analysis of Pan Kla ( H ) and H3K18la ( I ) protein levels in normal ( n = 30), OLK ( n = 30), and OSCC ( n = 30) tissues were assessed by immunohistochemical staining. Scale bar: 100 μm. ( J ) Pan Kla and H3K18la levels in human HOK, DOK, and OSCC cells (HSC3, HN4, HN6, and CAL27) were analyzed by western blotting assays. Error bars, mean ± SD; * P < 0.05.

Article Snippet: The human OSCC cell lines (HSC3, CAL27 and UPCI-SCC-154) were obtained from the American Type Culture Collection (VA, USA).

Techniques: Marker, Expressing, Immunohistochemical staining, Staining, Western Blot

Journal: Cell Death & Disease

Article Title: Histone lactylation-driven feedback loop modulates pyrimidine metabolism to promote oral carcinogenesis

doi: 10.1038/s41419-026-08580-w

Figure Lengend Snippet: A TK1 mRNA levels in normal ( n = 30) and OSCC ( n = 308) tissues were analyzed using the data from TCGA-OSCC cohort. B Kaplan–Meier survival analysis was performed to analyze overall survival based on TK1 mRNA levels (Low TK1 versus High TK1) in patients with OSCC in TCGA cohort ( n = 308). C TK1 mRNA levels in normal ( n = 30), OLK ( n = 30), and OSCC ( n = 30) tissues were measured using real-time RT-PCR. D – I DOK and CAL27 cells stably expressing NC or sh-TK1 were used to assess the effects of TK1 silencing on cell proliferation. The TK1 protein levels were measured using western blotting assays ( D ). Cell growth was evaluated using CCK-8 assay ( E ) and colony formation assay ( F , G ). The cell cycle distribution was measured using flow cytometry ( H , I ). Each experiment was performed in triplicate. Error bars, mean ± SD; * P < 0.05.

Article Snippet: The human OSCC cell lines (HSC3, CAL27 and UPCI-SCC-154) were obtained from the American Type Culture Collection (VA, USA).

Techniques: Quantitative RT-PCR, Stable Transfection, Expressing, Western Blot, CCK-8 Assay, Colony Assay, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: ALDH1L1 modulates CD8⁺ T-cell proliferation through the IL-15 signaling pathway. ( A ) Flow cytometric analysis of CD8⁺ T cell infiltration in clinical OSCC tissues ( n = 6 per group). ( B ) Expression distribution of ALDH1L1 across cell subtypes in OSCC from the TISCH2 database ( GSE172577 dataset). ( C ) Immunohistochemical staining of ALDH1L1 and CD8 in OSCC sections (ALDH1L1 low , n = 23; ALDH1L1 high , n = 29). ( D ) Immunohistochemical staining of ALDH1L1 and CD8 in xenograft tumor sections ( n = 6 per group). ( E ) Quantitative analysis of CD8⁺ T cell proportions in xenograft tissues by flow cytometry ( n = 6). ( F - G ) Correlation analysis between ALDH1L1 and chemokine mRNA expression levels in clinical OSCC samples (qRT-PCR, n = 12). H. qRT-PCR analysis of IL-15 mRNA expression in NC and ALDH1L1-KD groups of CAL 27 and SCC-25 cells ( n = 3). I. Extracellular IL-15 concentration in OSCC cell cultures with different ALDH1L1 expression levels ( n = 3). ( J ) Flow cytometric analysis of IL-15-mediated CD8⁺ T-cell proliferation in vitro, ( n = 3). (NC, negative control; OE, overexpression; KD, knockdown, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Immunohistochemical staining, Staining, Flow Cytometry, Quantitative RT-PCR, Concentration Assay, In Vitro, Negative Control, Over Expression, Knockdown

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: ALDH1L1 modulates CD8⁺ T-cell function through metabolic reprogramming. ( A ) Quantification of PD-1⁺ CD8⁺ T cells in clinical OSCC specimens by flow cytometry ( n = 6 per group). ( B ) Quantification of PD-1⁺ CD8⁺ T cells in mouse xenografts by flow cytometry ( n = 6 per group). ( C ) Top 20 enriched KEGG pathways identified from transcriptomic analysis. ( D ) Chord diagram illustrating 29 key metabolites identified by metabolomic profiling. ( E - H ) Flow cytometric evaluation of CD8⁺ T cell marker expression after stimulation with metabolites derived from ALDH1L1-KD or control CAL 27 cells and SCC-25 cells ( n = 3). (NC, negative control; OE, overexpression; KD, knockdown; ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Cell Function Assay, Flow Cytometry, Metabolomic, Marker, Expressing, Derivative Assay, Control, Negative Control, Over Expression, Knockdown

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: L-glutamate accumulation mediates CD8⁺ T cell dysfunction in ALDH1L1-downregulated microenvironment. ( A - C ) Integrated multi-omics analysis reveals ALDH1L1-regulated metabolic signatures: Red boxes highlight co-enriched metabolic pathways from transcriptomic ( A ) and metabolomic ( B ) analysis, and key metabolites ( C ). ( D ) Extracellular L-glutamate concentration in conditioned media from CAL 27 and SCC-25 cell culture supernatants ( n = 3). ( E - F ) Flow cytometric analysis of L-glutamate effects on CD8⁺ T cell functional markers: ( E ) Proportion of IFN-γ⁺ CD8⁺ T cells; ( F ) Proportion of PD-1⁺ CD8⁺ T cells ( n = 3). ( G ) Cytotoxic activity of CD8⁺ T cells pretreated with conditioned medium from ALDH1L1-KD or NC OSCC cells medium with exogenous L-glutamate ( n = 3). (NC, negative control; KD, knockdown; Ctrl, control; Glu, L-glutamate; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Biomarker Discovery, Metabolomic, Concentration Assay, Cell Culture, Functional Assay, Activity Assay, Negative Control, Knockdown, Control

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: ALDH1L1 transcriptionally regulates GLUL and interacts with GLUL to sustain L-glutamate metabolism. ( A ) Integrated multi-omics analysis of ALDH1L1-regulated metabolic network. Green circles: downregulated metabolites; Green squares: downregulated metabolic enzymes; Red squares: upregulated metabolic enzymes. ( B - C ) GLUL mRNA ( B ) and protein ( C ) expression in CAL 27 and SCC-25 cells assessed by qRT-PCR and western blot, respectively. ( D ) Protein-protein interaction (PPI) network between ALDH1L1 and glutamate-metabolizing enzymes predicted by the STRING database. ( E ) Three-dimensional structural model of ALDH1L1 (green)-GLUL (blue) interaction. Key residues are shown in stick representation; hydrogen bonds indicated by yellow dashed lines. ( F ) Co-immunoprecipitation analysis of endogenous interactions among ALDH1L1 and GLUL in CAL 27 cells. ( G ) ALDH1L1 enhances GLUL enzymatic activity in a dose-dependent manner in vitro ( n = 3). ( H ) Western blot validation of GLUL overexpression in plasmid-transfected CAL 27 cells. ( I ) Extracellular L-glutamate concentration following GLUL upregulation in ALDH1L1-KD CAL 27 cells ( n = 3). (NC: negative control; KD: ALDH1L1 knockdown; OE: GLUL overexpression; ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Western Blot, Immunoprecipitation, Activity Assay, In Vitro, Over Expression, Plasmid Preparation, Transfection, Concentration Assay, Negative Control, Knockdown

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: Stevioside enhances anti-PD-1 immunotherapy by targeting ALDH1L1. ( A ) Virtual screening workflow for identifying potential ALDH1L1-targeting compounds. ( B ) Two-dimensional chemical structures of the top 12 candidate compounds ranked by binding affinity to ALDH1L1 protein. ( C ) Western blot analysis demonstrating the effect of stevioside treatment on ALDH1L1 protein expression levels in CAL 27 cells (Ctrl, control). ( D ) Dose-response curve of stevioside (IC₅₀ = 195.3 µM). ( E ) Molecular docking analysis reveals the binding site of stevioside on ALDH1L1 protein. (Blue ribbon structure represents ALDH1L1 protein; green stick model represents stevioside molecule; stick structures indicate key interacting amino acid residues; yellow dashed lines denote hydrogen bond interactions.) ( F ) In vivo experiments confirm that stevioside combined with anti-PD-1 antibody treatment significantly enhances antitumor efficacy ( n = 5 per group). ( G - H ) Immunohistochemical staining of Ki-67 ( G ) and CD8 ( H ) in xenograft tumor sections ( n = 5 per group). (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Binding Assay, Western Blot, Expressing, Control, In Vivo, Immunohistochemical staining, Staining

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: Schematic model illustrating the mechanism by which ALDH1L1 reverses CD8⁺ T cell exhaustion in OSCC

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques:

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: ALDH1L1 modulates CD8⁺ T-cell proliferation through the IL-15 signaling pathway. ( A ) Flow cytometric analysis of CD8⁺ T cell infiltration in clinical OSCC tissues ( n = 6 per group). ( B ) Expression distribution of ALDH1L1 across cell subtypes in OSCC from the TISCH2 database ( GSE172577 dataset). ( C ) Immunohistochemical staining of ALDH1L1 and CD8 in OSCC sections (ALDH1L1 low , n = 23; ALDH1L1 high , n = 29). ( D ) Immunohistochemical staining of ALDH1L1 and CD8 in xenograft tumor sections ( n = 6 per group). ( E ) Quantitative analysis of CD8⁺ T cell proportions in xenograft tissues by flow cytometry ( n = 6). ( F - G ) Correlation analysis between ALDH1L1 and chemokine mRNA expression levels in clinical OSCC samples (qRT-PCR, n = 12). H. qRT-PCR analysis of IL-15 mRNA expression in NC and ALDH1L1-KD groups of CAL 27 and SCC-25 cells ( n = 3). I. Extracellular IL-15 concentration in OSCC cell cultures with different ALDH1L1 expression levels ( n = 3). ( J ) Flow cytometric analysis of IL-15-mediated CD8⁺ T-cell proliferation in vitro, ( n = 3). (NC, negative control; OE, overexpression; KD, knockdown, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Immunohistochemical staining, Staining, Flow Cytometry, Quantitative RT-PCR, Concentration Assay, In Vitro, Negative Control, Over Expression, Knockdown

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: ALDH1L1 modulates CD8⁺ T-cell function through metabolic reprogramming. ( A ) Quantification of PD-1⁺ CD8⁺ T cells in clinical OSCC specimens by flow cytometry ( n = 6 per group). ( B ) Quantification of PD-1⁺ CD8⁺ T cells in mouse xenografts by flow cytometry ( n = 6 per group). ( C ) Top 20 enriched KEGG pathways identified from transcriptomic analysis. ( D ) Chord diagram illustrating 29 key metabolites identified by metabolomic profiling. ( E - H ) Flow cytometric evaluation of CD8⁺ T cell marker expression after stimulation with metabolites derived from ALDH1L1-KD or control CAL 27 cells and SCC-25 cells ( n = 3). (NC, negative control; OE, overexpression; KD, knockdown; ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Cell Function Assay, Flow Cytometry, Metabolomic, Marker, Expressing, Derivative Assay, Control, Negative Control, Over Expression, Knockdown

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: L-glutamate accumulation mediates CD8⁺ T cell dysfunction in ALDH1L1-downregulated microenvironment. ( A - C ) Integrated multi-omics analysis reveals ALDH1L1-regulated metabolic signatures: Red boxes highlight co-enriched metabolic pathways from transcriptomic ( A ) and metabolomic ( B ) analysis, and key metabolites ( C ). ( D ) Extracellular L-glutamate concentration in conditioned media from CAL 27 and SCC-25 cell culture supernatants ( n = 3). ( E - F ) Flow cytometric analysis of L-glutamate effects on CD8⁺ T cell functional markers: ( E ) Proportion of IFN-γ⁺ CD8⁺ T cells; ( F ) Proportion of PD-1⁺ CD8⁺ T cells ( n = 3). ( G ) Cytotoxic activity of CD8⁺ T cells pretreated with conditioned medium from ALDH1L1-KD or NC OSCC cells medium with exogenous L-glutamate ( n = 3). (NC, negative control; KD, knockdown; Ctrl, control; Glu, L-glutamate; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Biomarker Discovery, Metabolomic, Concentration Assay, Cell Culture, Functional Assay, Activity Assay, Negative Control, Knockdown, Control

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: ALDH1L1 transcriptionally regulates GLUL and interacts with GLUL to sustain L-glutamate metabolism. ( A ) Integrated multi-omics analysis of ALDH1L1-regulated metabolic network. Green circles: downregulated metabolites; Green squares: downregulated metabolic enzymes; Red squares: upregulated metabolic enzymes. ( B - C ) GLUL mRNA ( B ) and protein ( C ) expression in CAL 27 and SCC-25 cells assessed by qRT-PCR and western blot, respectively. ( D ) Protein-protein interaction (PPI) network between ALDH1L1 and glutamate-metabolizing enzymes predicted by the STRING database. ( E ) Three-dimensional structural model of ALDH1L1 (green)-GLUL (blue) interaction. Key residues are shown in stick representation; hydrogen bonds indicated by yellow dashed lines. ( F ) Co-immunoprecipitation analysis of endogenous interactions among ALDH1L1 and GLUL in CAL 27 cells. ( G ) ALDH1L1 enhances GLUL enzymatic activity in a dose-dependent manner in vitro ( n = 3). ( H ) Western blot validation of GLUL overexpression in plasmid-transfected CAL 27 cells. ( I ) Extracellular L-glutamate concentration following GLUL upregulation in ALDH1L1-KD CAL 27 cells ( n = 3). (NC: negative control; KD: ALDH1L1 knockdown; OE: GLUL overexpression; ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Western Blot, Immunoprecipitation, Activity Assay, In Vitro, Over Expression, Plasmid Preparation, Transfection, Concentration Assay, Negative Control, Knockdown

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: Stevioside enhances anti-PD-1 immunotherapy by targeting ALDH1L1. ( A ) Virtual screening workflow for identifying potential ALDH1L1-targeting compounds. ( B ) Two-dimensional chemical structures of the top 12 candidate compounds ranked by binding affinity to ALDH1L1 protein. ( C ) Western blot analysis demonstrating the effect of stevioside treatment on ALDH1L1 protein expression levels in CAL 27 cells (Ctrl, control). ( D ) Dose-response curve of stevioside (IC₅₀ = 195.3 µM). ( E ) Molecular docking analysis reveals the binding site of stevioside on ALDH1L1 protein. (Blue ribbon structure represents ALDH1L1 protein; green stick model represents stevioside molecule; stick structures indicate key interacting amino acid residues; yellow dashed lines denote hydrogen bond interactions.) ( F ) In vivo experiments confirm that stevioside combined with anti-PD-1 antibody treatment significantly enhances antitumor efficacy ( n = 5 per group). ( G - H ) Immunohistochemical staining of Ki-67 ( G ) and CD8 ( H ) in xenograft tumor sections ( n = 5 per group). (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Binding Assay, Western Blot, Expressing, Control, In Vivo, Immunohistochemical staining, Staining

Journal: Journal of Translational Medicine

Article Title: ALDH1L1 reverses CD8 + T cell exhaustion in the oral squamous cell carcinoma microenvironment by reprogramming L-glutamate metabolism

doi: 10.1186/s12967-026-07812-z

Figure Lengend Snippet: Schematic model illustrating the mechanism by which ALDH1L1 reverses CD8⁺ T cell exhaustion in OSCC

Article Snippet: The human OSCC cell lines CAL 27 and SCC-25 were purchased from Procell Life Science & Technology Co., Ltd. (China), while the murine OSCC cell line SCC-7 was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: