Journal: BMC Oral Health

Article Title: Targeting CGRP signaling alleviates cancer-associated pain in oral squamous cell carcinoma

doi: 10.1186/s12903-026-08444-x

Figure Lengend Snippet: Analgesic effects of Rimegepant in OSCC-associated pain demonstrated via the unilateral tongue orthotopic transplantation model. A Construction of the unilateral tongue orthotopic transplantation model and drug injection with mechanical threshold measurement protocol. B Following tumor transplantation, the differences in mechanical sensitivity between the transplanted ( n = 14) and non-transplanted ( n = 10) sides were compared using unpaired two-tailed Student’s t test. C Changes in sensitivity 1 h after Rimegepant (50 mg/kg), Rimegepant (10 mg/kg) and Carprofen treatment were performed using one-way ANOVA. Each dataset included results from monitoring over 4 consecutive days ( n = 6–8/group). D Changes in sensitivity following Rimegepant (50 mg/kg) and Tramadol treatment at 1 h were performed using one-way ANOVA. Each dataset included results from monitoring over 4 consecutive days ( n = 6–8/group). E The 4-day trends of facial mechanical sensitivities 24 h after Rimegepant (50 mg/kg) and Tramadol treatment. ( n = 6–8/group). F Changes in sensitivity 1 h after Tramadol, Carprofen treatment and local lingual injection of Rimegepant (10 mg/kg) in CAL27 and SCC25 xenograft models were performed using one-way ANOVA( n = 8). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns P > 0.05

Article Snippet: Human OSCC CAL27 and SCC25 cells were purchased from the American Type Culture Collection (USA).

Techniques: Transplantation Assay, Injection, Two Tailed Test

Journal: Cancer Cell International

Article Title: LIS1 mediated Schwann cell reprogramming enhances perineural invasion by activating the serine/NMDAR/AKT signaling pathway in head and neck squamous carcinoma

doi: 10.1186/s12935-026-04243-0

Figure Lengend Snippet: LIS1 increases the migration, invasion, and PNI capability of HNSCC cells. ( A ) Wound healing assay of LIS1 overexpression or LIS1 knockdown in SCC25 cells. ( B ) Transwell assay of LIS1 overexpression or LIS1 knockdown in SCC25 cells. ( C ) Representative images of the in vitro PNI model for LIS1 overexpression or LIS1 knockdown in SCC25 cells. ( D ) Microfluidic chip pattern diagram and representative images of LIS1 overexpression or LIS1 knockdown in SCC25 cells co-cultured with SCs in the microfluidic chip. Data are represented as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human HNSCC cell line (SCC25) and human Schwann cells (sNF96.2) were acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA) in 2023.

Techniques: Migration, Wound Healing Assay, Over Expression, Knockdown, Transwell Assay, In Vitro, Cell Culture

Journal: Cancer Cell International

Article Title: LIS1 mediated Schwann cell reprogramming enhances perineural invasion by activating the serine/NMDAR/AKT signaling pathway in head and neck squamous carcinoma

doi: 10.1186/s12935-026-04243-0

Figure Lengend Snippet: LIS1 overexpression of HNSCC cells increases Schwann cells activity. ( A ) The pattern diagram of HNSCC cells co-cultured with SCs. ( B ) Transwell assay of sNF96.2 cells co-cultured with SCC25 cells. ( C ) The mRNA expression of PNI-related factors (BDNF, GDNF, NGF, MMP2, and MMP9) in sNF96.2 cells co-cultured with SCC25 cells were analyzed by RT-qPCR. ( D ) IF double staining was performed to detect the expression of S100 and GFAP in the sNF96.2 cells co-cultured with SCC25 cells. Data are represented as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human HNSCC cell line (SCC25) and human Schwann cells (sNF96.2) were acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA) in 2023.

Techniques: Over Expression, Activity Assay, Cell Culture, Transwell Assay, Expressing, Quantitative RT-PCR, Double Staining

Journal: Cancer Cell International

Article Title: LIS1 mediated Schwann cell reprogramming enhances perineural invasion by activating the serine/NMDAR/AKT signaling pathway in head and neck squamous carcinoma

doi: 10.1186/s12935-026-04243-0

Figure Lengend Snippet: LIS1 overexpression activates the serine signaling pathway to promote the PNI in HNSCC cells. ( A ) The mRNA expression of PHGDH, PSAT1, and PSPH in SCC25 cells were analyzed by RT-qPCR. ( B ) Analysis of the relative intracellular serine levels in SCC25 cells. ( C ) The protein level of PHGDH, PSAT1, and PSPH in SCC25 cells were analyzed by WB. ( D ) Migration and invasion assay of shLIS1 group after exogenous serine stimulation of SCC25 cells and quantitative analysis. ( E ) Representative images of the in vitro PNI model for shLIS1 group after exogenous serine stimulation of SCC25 cells and the analysis of neural invasion index. ( F ) Representative images of the microfluidic chip for shLIS1 group after exogenous serine stimulation of SCC25 cells and analysis of migration distance. Data are represented as mean ± SEM, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human HNSCC cell line (SCC25) and human Schwann cells (sNF96.2) were acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA) in 2023.

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Migration, Invasion Assay, In Vitro

Journal: Journal of Dental Sciences

Article Title: Tumor microenvironment-derived IL-32 promotes aggressive phenotypes and stem cell traits in head and neck squamous cell carcinoma

doi: 10.1016/j.jds.2025.10.010

Figure Lengend Snippet: Interleukin-32 (IL-32) induces EMT in HNSCC cells, and neutralisation of IL-32 attenuates EMT-associated morphology and proteins. Phase-contrast images of FaDu and SCC25 cells cultured for 24 h in control medium, CAF-conditioned medium (CAF-CM), recombinant IL-32 (rIL-32, 50 ng mL −1 ), or CAF-CM plus an IL-32-neutralising antibody (IL-32 Ab). CAF-CM and rIL-32 induce elongated, spindle-like morphology with microspike-like protrusions (red arrows), whereas addition of IL-32 Ab markedly suppresses these features. Scale bars = 200 μm (A). Immunoblots showing EMT-related protein changes in FaDu and SCC25 cells with or without rIL-32. rIL-32 increases IL-32, Snail, Twist, Vimentin and Fibronectin, while reducing the epithelial marker E-cadherin. β-Tubulin serves as the loading control (B). Western-blot comparison of CAF-CM versus CAF-CM + IL-32 Ab. Neutralising IL-32 diminishes IL-32, Snail and Twist, restores E-cadherin, and lowers Vimentin and Fibronectin levels (C). Together these data confirm that IL-32 is sufficient to trigger EMT in head and neck squamous cell carcinoma cells.

Article Snippet: FaDu (hypopharyngeal) and SCC25 (tongue) HNSCC cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Cell Culture, Control, Recombinant, Western Blot, Marker, Comparison

Journal: Journal of Dental Sciences

Article Title: Tumor microenvironment-derived IL-32 promotes aggressive phenotypes and stem cell traits in head and neck squamous cell carcinoma

doi: 10.1016/j.jds.2025.10.010

Figure Lengend Snippet: CAF-derived interleukin-32 (IL-32) promotes migration and invasion of HNSCC cells, and IL-32 neutralisation reverses these effects. Migration assay. Crystal-violet-stained FaDu and SCC25 cells that traversed 8 μm Transwell filters after 24 h in control medium, CAF-CM or recombinant IL-32 (50 ng mL −1 ) or CAF-CM plus an IL-32-neutralising antibody (CAF-CM + IL-32 Ab). CAF-CM and rIL-32 markedly increased motility, whereas addition of IL-32 Ab reduced migration towards near-baseline levels. Right, quantification as fold change relative to control (mean ± SD, n = 3; one-way ANOVA with Tukey's post-hoc; ∗∗ P < 0.01 vs control). Scale bars = 1 mm (A). Invasion assay. Cells were seeded on Matrigel-coated inserts and allowed to invade for 48 h under the same conditions. Quantification (right) indicates a significant 1.5- to 2-fold increase in invading cells with CAF-CM or rIL-32, while IL-32 Ab blunted this effect (mean ± SD, n = 3; ∗∗ P < 0.01 vs control). Scale bars = 1 mm (B). Three-dimensional organotypic model. Haematoxylin-and-eosin-stained sections of collagen/Matrigel gels populated with normal fibroblasts (NF) or CAFs and overlaid with FaDu or SCC25 cells. After 14 d, CAF co-culture produced deeper epithelial invasion (arrows) compared with NF controls, whereas inclusion of IL-32 Ab with CAF-CM largely abolished invasion. Scale bars = 1 mm (C). Collectively, these data confirm that CAF-secreted IL-32 potentiates HNSCC cell migration and invasion in both 2-D and physiologically relevant 3-D settings.

Article Snippet: FaDu (hypopharyngeal) and SCC25 (tongue) HNSCC cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Derivative Assay, Migration, Staining, Control, Recombinant, Invasion Assay, Co-Culture Assay, Produced

Journal: Journal of Dental Sciences

Article Title: Tumor microenvironment-derived IL-32 promotes aggressive phenotypes and stem cell traits in head and neck squamous cell carcinoma

doi: 10.1016/j.jds.2025.10.010

Figure Lengend Snippet: Interleukin-32 (IL-32) enhances cancer-stem-cell (CSC) properties in HNSCC cells. Representative phase-contrast micrographs of tumourspheres formed by FaDu and SCC25 cells cultured for 10 days in serum-free medium with or without recombinant IL-32 (rIL-32, 50 ng mL −1 ). rIL-32 treatment markedly increases both sphere size and number. Scale bars = 1 mm (A). Immunoblots showing stemness-associated transcription factors Oct-4, SOX2 and NANOG in control (WT) and rIL-32-treated cells. β-Tubulin serves as a loading control; rIL-32 elevates all three markers, indicating augmented stemness (B). Flow-cytometric analysis of surface CSC markers. Histograms illustrate CD24, CD44 and CD133 expression profiles for WT and rIL-32-treated FaDu (left) and SCC25 (right) populations; M2 gates (purple) represent positive cells. Bar graphs (bottom) depict mean ± SD fold change in CD44 + /CD24 + and CD133 + subsets (n = 3). ∗ P < 0.05; ∗∗∗ P < 0.001 versus control (two-tailed t-test). These data demonstrate that IL-32 not only drives EMT and invasion but also potentiates CSC traits in head and neck squamous cell carcinoma cells (C).

Article Snippet: FaDu (hypopharyngeal) and SCC25 (tongue) HNSCC cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Cell Culture, Recombinant, Western Blot, Control, Expressing, Two Tailed Test