Journal: Antioxidants

Article Title: Saposhnikovia divaricata Inhibits Inflammation, Oxidative Stress, and Ferroptosis to Alleviate DSS-Induced Ulcerative Colitis

doi: 10.3390/antiox15020258

Figure Lengend Snippet: FF modulates the expression of ferroptosis-related proteins in colon tissue via the p53 pathway. ( A ) Representative immunohistochemical (IHC) images of p53, SLC7A11, and GPX4 expression in colon sections (scale bar = 50 μm). ( B – D ) Quantitative analysis of the relative protein expression levels of p53 (B), SLC7A11 (C), and GPX4 (D). Data are presented as the mean ± SD ( n = 3 independent experiments). ### p < 0.001 versus the control (CON) group; * p < 0.05, ** p < 0.01, *** p < 0.001 versus the DSS model group.

Article Snippet: Primary antibodies against GPX4 (Item #67763-1-Ig) and SLC7A11 (Item #26864-1-AP) were purchased from Proteintech Group Inc. (Chicago, IL, USA).

Techniques: Expressing, Immunohistochemical staining, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Sulfasalazine combined with anti-IL-1β mAb induces ferroptosis and immune modulation in oral squamous cell carcinoma

doi: 10.1007/s00018-025-05742-5

Figure Lengend Snippet: High expression of SLC7A11 correlates with poor prognosis in OSCC patients. ( A ) The differential expression between tumor and normal tissues for SLC7A11 across all TCGA tumors analyzed by TIMER2.0. ( B ) The expression of SLC7A11 in adjacent tissues and OSCC tissues from the TCGA database. ( C ) Representative IHC images (left) and quantitative analysis (right) of SLC7A11 expression in adjacent tissues and OSCC tissues from patients. Scale bar: 200 µm for 10 × magnification and 40 µm for 40 × magnification. ( D ) Western blot analysis of SLC7A11 expression levels in OSCC cell lines compared with NOK. ( E ) Kaplan-Meier survival analysis of SLC7A11 expression in HNSCC patients by using GEPIA. Values are presented as mean ± SEM. In A, Wilcoxon test; B, Mann-Whitney U test; C, unpaired Student’s t-test; D, one-way ANOVA; E, Log-rank test

Article Snippet: Briefly, the sections of 4NQO-rat tongues were incubated with primary antibodies against SLC7A11 (1:100, DF12509, Affinity Biosciences), PTGS2 (1:200, 66351, Proteintech, USA), IL-1β (1:500, ab283818, Abcam, USA), Ki67 (1:200, GB151499 , Servicebio, China) at 4 °C overnight.

Techniques: Expressing, Quantitative Proteomics, Western Blot, MANN-WHITNEY

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Sulfasalazine combined with anti-IL-1β mAb induces ferroptosis and immune modulation in oral squamous cell carcinoma

doi: 10.1007/s00018-025-05742-5

Figure Lengend Snippet: SAS suppresses OSCC cells proliferation via inducing ferroptosis in vitro. ( A ) Dose-response for SCC1 after exposure to indicated concentrations of SAS for 48 h. IC50 concentrations were listed and error bars indicated SEM for quadruplicate measurements. ( B ) Cell viability of SCC1 after 800 μM SAS treatment by CCK-8. The differences were considered statistically significant if (*) P < 0.05, (**) P < 0.01, (***) P < 0.001, (****) P < 0.0001. ( C ) Cell proliferation of SCC1 after 800 µM SAS treatment for 24 h quantified by EdU assay. Scale bar = 100 μm. ( D ) Intracellular ROS levels of SCC1 after 800 µM SAS treatment for 24 h measured by DCFH-DA staining via flow cytometry. ( E ) Intracellular GSH levels of SCC1 after 800 µM SAS treatment for 24 h measured by GSH Quantification Kit. ( F ) Lipid peroxidation of SCC1 after 800 µM SAS treatment for 24 h detected by BODIPY 581/591 C11. Scale bar = 100 µm. ( G ) Q-PCR experiment of SLC7A11 and PTGS2 expressions in SCC1 after 800 µM SAS inducing for 24 h. (H) Quantitative analysis of WB for SLC7A11 and PTGS2 expressions in SCC1 after 800 µM SAS inducing for 24 h. ( I ) TEM images of SCC1 cells after 800 µM SAS treatment for 24 h with analyzed relative mitochondrial size and percentage of damaged mitochondria. Scale bars: 5 µm and 500 nm. Values are presented as mean ± SEM. In B-I, unpaired Student’s t-test

Article Snippet: Briefly, the sections of 4NQO-rat tongues were incubated with primary antibodies against SLC7A11 (1:100, DF12509, Affinity Biosciences), PTGS2 (1:200, 66351, Proteintech, USA), IL-1β (1:500, ab283818, Abcam, USA), Ki67 (1:200, GB151499 , Servicebio, China) at 4 °C overnight.

Techniques: In Vitro, CCK-8 Assay, IF-P, EdU Assay, Staining, Flow Cytometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Sulfasalazine combined with anti-IL-1β mAb induces ferroptosis and immune modulation in oral squamous cell carcinoma

doi: 10.1007/s00018-025-05742-5

Figure Lengend Snippet: SAS-induced accumulation of IL-1β correlates with T cell exhaustion. ( A ) Schematic view of SAS CM and T cells isolated from PBMCs. ( B ) The levels of IL-8, IL-1β, IL-6, IL-10, TNF-α and IL-12p70 in the SAS CM analyzed by CBA. ( C ) Q-PCR experiment of IL-1β expression in SCC1 after SAS (800 µM) inducing for 24 h. ( D ) Quantitative analysis of WB for IL-1β expression in SCC1 after SAS (800 µM) inducing for 24 h. ( E ) The level of IL-1β in the SAS CM measured by ELISA. ( F ) Flow cytometric analysis detecting the levels of PD-1, CTLA-4, TNF-α and IFN-γ of 40% SAS CM treating CD8+T cells. ( G ) Flow cytometric analysis detecting the levels of PD-1, CTLA-4, TNF-α and IFN-γ of CD8+T cells treated by 30 pg/mL IL-1β for 24 h. ( H ) The expression of IL-1β in adjacent tissues and OSCC tissues from the TCGA database. ( I ) The correlation between CD8+ T cell infiltration and IL-1β expression for OSCC patients in the TCGA database. ( J ) Kaplan-Meir survival analysis of SLC7A11 expression for OSCC patients in the TCGA database. Values are presented as mean ± SEM. In B, unpaired Student’s t-test or Mann-Whitney U test; C, D, E, F, G and H, unpaired Student’s t-test; J, Log-rank test

Article Snippet: Briefly, the sections of 4NQO-rat tongues were incubated with primary antibodies against SLC7A11 (1:100, DF12509, Affinity Biosciences), PTGS2 (1:200, 66351, Proteintech, USA), IL-1β (1:500, ab283818, Abcam, USA), Ki67 (1:200, GB151499 , Servicebio, China) at 4 °C overnight.

Techniques: Isolation, Expressing, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Sulfasalazine combined with anti-IL-1β mAb induces ferroptosis and immune modulation in oral squamous cell carcinoma

doi: 10.1007/s00018-025-05742-5

Figure Lengend Snippet: Combination of SAS and anti-IL-1β mAb enhances ferroptosis and inhibits OSCC cell growth. ( A ) Cell viability of SCC1, A375 and MDA-MB231 after 800 μM SAS or/and 50 pM anti-IL-1β mAb treatment by CCK-8. ( B ) A diagram of Jurkat T cells and cancer cells co-culture model. ( C ) Live cells of SCC1, A375 and MDA-MB231 after 800 μM SAS or/and 50 pM anti-IL-1β mAb treatment and co-culture with Jurkat T cells for 24 h. ( D ) EdU assay to quantify cell proliferation of SCC1 after 800 μM SAS or/and 50 pM anti-IL-1β mAb treatment for 24 h. Scale bar = 100 μm. ( E ) Intracellular ROS levels measured by DCFH-DA staining via flow cytometry. SCC1 cells were incubated with 800 μM SAS or/and 50 Pm anti-IL-1β mAb treatment for 24 h. ( F ) Intracellular GSH of SCC1 measured by a GSH Quantification Kit after 800 μM SAS or/and 50 pM anti-IL-1β mAb treatment for 24 h. ( G ) Lipid peroxidation of SCC1 detected by BODIPY 581/591 C11 after 800 μM SAS or/and 50 pM anti-IL-1β mAb treatment for 24 h. Scale bar = 100 μm. ( H ) Q-PCR experiment of SLC7A11 and PTGS2 expressions in SCC1 after 800 μM SAS or/and 50 pM anti-IL-1β mAb inducing for 24 h. ( I ) Quantitative analysis of WB for SLC7A11 and PTGS2 expressions in SCC1 after 800 μM SAS or/and 50 pM anti-IL-1β mAb inducing for 24 h. Values are presented as mean ± SEM. In A, E, F, G, H and I, one-way ANOVA; C, unpaired Student’s t-test; D, Kruskal-Wallis test

Article Snippet: Briefly, the sections of 4NQO-rat tongues were incubated with primary antibodies against SLC7A11 (1:100, DF12509, Affinity Biosciences), PTGS2 (1:200, 66351, Proteintech, USA), IL-1β (1:500, ab283818, Abcam, USA), Ki67 (1:200, GB151499 , Servicebio, China) at 4 °C overnight.

Techniques: CCK-8 Assay, Co-Culture Assay, EdU Assay, Staining, Flow Cytometry, Incubation

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Sulfasalazine combined with anti-IL-1β mAb induces ferroptosis and immune modulation in oral squamous cell carcinoma

doi: 10.1007/s00018-025-05742-5

Figure Lengend Snippet: Combination of SAS and anti-IL-1β mAb inhibits oral carcinogenesis in 4NQO induced rat model. ( A ) Schematic overview of the 4NQO model experimental design. ( B ) Gross observation of the rat tongues in different groups at the endpoint. Scale bars: 1 cm for gross. ( C ) Quantification of the histological degree of moderate dysplasia, severe dysplasia, and carcinoma in situ and invasive carcinoma in the four groups. ( D ) H&E scores of the histopathologic diagnoses in the four groups. ( E ) Representative H&E images of the rat tongues in different groups at the endpoint. Scale bars: 300 µm for 10 × magnification. ( F ) Representative IHC staining of SLC7A11, PTGS2, IL-1β and Ki67. Data points represent individual rats (n = 6 per group). Scale bars: 50 µm. Values are presented as means ± SEM. In D and F, one-way ANOVA

Article Snippet: Briefly, the sections of 4NQO-rat tongues were incubated with primary antibodies against SLC7A11 (1:100, DF12509, Affinity Biosciences), PTGS2 (1:200, 66351, Proteintech, USA), IL-1β (1:500, ab283818, Abcam, USA), Ki67 (1:200, GB151499 , Servicebio, China) at 4 °C overnight.

Techniques: In Situ, Immunohistochemistry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Sulfasalazine combined with anti-IL-1β mAb induces ferroptosis and immune modulation in oral squamous cell carcinoma

doi: 10.1007/s00018-025-05742-5

Figure Lengend Snippet: Schematic model of the combination effect of SAS with anti-IL-1β mAb. ( A ) SLC7A11 is a crucial transmembrane transporter involved in the extracellular uptake of cystine. Cystine enters cells via SLC7A11, facilitating the synthesis of the potent antioxidant GSH, which mitigates ROS accumulation, prevents lipid peroxidation, and reduces tumor cell death. Within the tumor microenvironment, T cells are present to target and eliminate tumor cells. ( B ) SAS acts as an inhibitor of SLC7A11. SLC7A11 inhibition decreases intracellular GSH levels, and increases ROS production and subsequent lipid peroxidation. Concurrently, the expression of PTGS2 rises, inducing ferroptosis in tumor cells. As ferroptosis progresses, a notable increase in IL-1β secretion is observed within the tumor microenvironment, resulting in T cell exhaustion. ( C ) This T cell dysfunction can be reversed by the addition of anti-IL-1β mAb. Combined therapy utilizing SAS and anti-IL-1β mAb effectively targets the vulnerability of iron-dependent cell death, thereby enhancing immune responses and promoting tumor cell destruction

Article Snippet: Briefly, the sections of 4NQO-rat tongues were incubated with primary antibodies against SLC7A11 (1:100, DF12509, Affinity Biosciences), PTGS2 (1:200, 66351, Proteintech, USA), IL-1β (1:500, ab283818, Abcam, USA), Ki67 (1:200, GB151499 , Servicebio, China) at 4 °C overnight.

Techniques: Inhibition, Expressing