human pharyngeal carcinoma pleural fluid metastatic cell detroit 562 cells  (ATCC)

Journal: iScience

Article Title: THBS1 upregulation by KLF7 in hypopharyngeal squamous cell carcinoma contributes to lung metastasis through the p38 MAPK signaling

doi: 10.1016/j.isci.2026.114816

Figure Lengend Snippet: THBS1 is highly expressed in HPSCC tissues and cell lines (A) A volcano plot demonstrating DEGs between HPSCC and normal control tissues in the GSE2379 dataset (adj. p Val <0.05, |LogFC| > 2). (B) The intersection of DEGs in the GSE2379 dataset and prognostic markers in HNSC. (C) The PPI protein interaction network of 42 intersecting genes by STRING (disconnected nodes in the work are not shown). (D) p -value and LogFC value of the THBS1 gene in the GSE2379 dataset. (E–G) Prognostic significance of THBS1 expression in patients with HNSC analyzed by UALCAN. RT-qPCR (F) and Western blot (G) analysis of THBS1 expression differences between tumors and paired peritumoral tissues in 22 patients with HPSCC. RT-qPCR (H) and Western blot (I) analysis of THBS1 expression differences between HOK, FaDu, and Detroit 562 cells. The results are expressed as the mean ± SD. (H–I). These values are derived from three independent experiments. Statistical analysis was performed using paired t test (F-G) or one-way ANOVA (H-I), followed by Tukey’s multiple comparison test.

Article Snippet: The HPSCC cell lines FaDu (CL-0083) and Detroit 562 (CL-0330) cells were purchased from Procell (Wuhan, Hubei, China).

Techniques: Control, Expressing, Quantitative RT-PCR, Western Blot, Derivative Assay, Comparison

Journal: iScience

Article Title: THBS1 upregulation by KLF7 in hypopharyngeal squamous cell carcinoma contributes to lung metastasis through the p38 MAPK signaling

doi: 10.1016/j.isci.2026.114816

Figure Lengend Snippet: Exogenous inhibition of THBS1 significantly curtails the malignant behavioral capacity of HPSCC cells. RT-qPCR (A and B) Western blot (B) analysis of THBS1 expression in FaDu and Detroit 562 cells after infection with KD-THBS1 #1–3. (C) HPSCC cell proliferative capacity was examined using EdU staining. (D) The colony formation of HPSCC cells. (E–I) The migratory capacity of HPSCC cells was examined using wound healing assays. Transwell assay detects changes in the migratory (F) and invasive (G) abilities of HPSCC cells. Expression of E-cadherin and N-cadherin in HPSCC cells was examined using Western blot analysis (H) and dual-labeling immunofluorescence (I). The results are expressed as the mean ± SD (A–I). These values are derived from three independent experiments. Statistical analysis was performed using a one-way ANOVA (A–I), followed by Tukey’s multiple comparison test.

Article Snippet: The HPSCC cell lines FaDu (CL-0083) and Detroit 562 (CL-0330) cells were purchased from Procell (Wuhan, Hubei, China).

Techniques: Inhibition, Quantitative RT-PCR, Western Blot, Expressing, Infection, Staining, Transwell Assay, Labeling, Immunofluorescence, Derivative Assay, Comparison

Journal: iScience

Article Title: THBS1 upregulation by KLF7 in hypopharyngeal squamous cell carcinoma contributes to lung metastasis through the p38 MAPK signaling

doi: 10.1016/j.isci.2026.114816

Figure Lengend Snippet: KLF7 transcriptional activation of THBS1 in HPSCC cells. RT-qPCR (A–D) Western blot (B) analysis of KLF7 expression differences between tumors and paired peritumoral tissues in 22 patients with HPSCC. RT-qPCR (C) and Western blot (D) analysis of KLF7 expression differences between HOK, FaDu, and Detroit 562 cells. (E and G) The degree of enrichment of the THBS1 promoter by anti-KLF7 in HPSCC cells was examined using ChIP. RT-qPCR (F) and Western blot analysis (G) of the knockdown efficiency of KLF7 in HPSCC cells. (H) The mRNA levels of THBS1 in knockdown KLF7-treated HPSCC cells were detected by RT-qPCR. (I) The luciferase activity of the THBS1 promoter in knockdown KLF7-treated HPSCC cells was detected by the dual-luciferase assay. (J) The luciferase activity of the THBS1 promoter in HPSCC cells transfected with WT or Mut vectors was detected by the dual-luciferase assay. The results are expressed as the mean ± SD (A–J). These values are derived from three independent experiments. Statistical analysis was performed using a paired t test (A–B), unpaired t test (E, H–I), one-way ANOVA (C–D, F-G), or two-way ANOVA (J), followed by Tukey’s multiple comparison test.

Article Snippet: The HPSCC cell lines FaDu (CL-0083) and Detroit 562 (CL-0330) cells were purchased from Procell (Wuhan, Hubei, China).

Techniques: Activation Assay, Quantitative RT-PCR, Western Blot, Expressing, Knockdown, Luciferase, Activity Assay, Transfection, Derivative Assay, Comparison

Journal: iScience

Article Title: THBS1 upregulation by KLF7 in hypopharyngeal squamous cell carcinoma contributes to lung metastasis through the p38 MAPK signaling

doi: 10.1016/j.isci.2026.114816

Figure Lengend Snippet: HDAC6 removes H3K9ac modification at the KLF7 promoter to inhibit KLF7 expression (A and B) Western blot analysis of HDAC6 expression differences between tumors and paired peritumoral tissues in 22 patients with HPSCC (A) and between HOK, FaDu, and Detroit 562 cells (B). (C and D) The degree of enrichment of the KLF7 promoter by anti-HDAC6 and anti-H3K9ac in HPSCC cells was examined using ChIP. Western blot analysis (D) of the overexpression efficiency of HDAC6 in HPSCC cells. (E) The degree of enrichment of the KLF7 promoter by anti-HDAC6 and anti-H3K9ac in HPSCC cells overexpressing HDAC6 was examined using ChIP. (F) KLF7 and THBS1 mRNA levels in HPSCC cells overexpressing HDAC6 were examined using RT-qPCR. (G) The binding of anti-H3K14ac and anti-H4K8ac to the KLF7 promoter in HPSCC cells overexpressing HDAC6 was examined using ChIP. The results are expressed as the mean ± SD (A–G). These values are derived from three independent experiments. Statistical analysis was performed using a paired test (A), unpaired t test (D, F), one-way ANOVA (B–C), and two-way ANOVA (E, G), followed by Tukey’s multiple comparison test.

Article Snippet: The HPSCC cell lines FaDu (CL-0083) and Detroit 562 (CL-0330) cells were purchased from Procell (Wuhan, Hubei, China).

Techniques: Modification, Expressing, Western Blot, Over Expression, Quantitative RT-PCR, Binding Assay, Derivative Assay, Comparison

Journal: PLOS One

Article Title: Context-dependent dysregulation of store-operated calcium channels in head and neck squamous cell carcinoma

doi: 10.1371/journal.pone.0344393

Figure Lengend Snippet: (A) Boxplot representation of mRNA expression levels of ORAI1, ORAI2, ORAI3, STIM1, and STIM2 across normal solid tissue (red), primary tumors (blue), and metastatic tissues (orange). Data were obtained from the UCSC Xena Browser using the TCGA HNSCC cohort (n = 566). Expression is shown as log₂(normalized count + 1). One-way ANOVA revealed significant differences across groups: ORAI1 (p < 0.001), ORAI2 (p < 0.001), ORAI3 (p < 0.01), STIM1 (p < 0.001), and STIM2 (p < 0.001). (B) Dot plot of ORAI and STIM isoform expression across HNSCC anatomical subsites in the TCGA cohort (n = 604), t-test (ORAI1: p > 0.05; ORAI2: p < 0.01; ORAI3: p < 0.01; STIM1: p < 0.05; STIM2: p > 0.05). (C) OncoPrint visualization of somatic alterations in ORAI/STIM genes in the TCGA PanCancer Atlas HNSCC cohort (n = 523). (D) OncoPrint visualization and heatmap of the same cohort showing mRNA alterations relative to normal samples.

Article Snippet: The HNSCC cell lines FaDu and Detroit-562 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing

Journal: PLOS One

Article Title: Context-dependent dysregulation of store-operated calcium channels in head and neck squamous cell carcinoma

doi: 10.1371/journal.pone.0344393

Figure Lengend Snippet: (A, B) Copy number alterations (A) and mRNA expression levels (B) of ORAI1, ORAI2, ORAI3, STIM1, and STIM2 in FaDu and Detroit-562 HNSCC cell lines, retrieved from the Cancer Cell Line Encyclopedia (CCLE) using the UCSC Xena browser. Heatmaps illustrate relative gene copy number (A) and transcript abundance (B) . (C) Quantitative RT-PCR validation of ORAI and STIM isoform expression in FaDu and Detroit-562 cells (n = 3). Expression was normalized to GAPDH, and relative levels were calculated using the 2 (-ΔCt) method. Data represent mean ± SEM. (D) EGFR mRNA expression (RPKM) in FaDu and Detroit-562 cells, retrieved from CCLE/Xena. Values represent summary statistics; error bars are not shown.

Article Snippet: The HNSCC cell lines FaDu and Detroit-562 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR, Biomarker Discovery

Journal: PLOS One

Article Title: Context-dependent dysregulation of store-operated calcium channels in head and neck squamous cell carcinoma

doi: 10.1371/journal.pone.0344393

Figure Lengend Snippet: (A, B) Dose–response curves of FaDu (A) and Detroit-562 (B) cell lines treated with increasing concentrations of 2-APB for 48 hours. Cell viability was assessed using the MTT assay. Viability was normalized to vehicle-treated control, and IC₅₀ values were calculated using non-linear regression (log[2-APB] vs. % viability) in GraphPad Prism 8.0. Data represent the mean ± SD of at least three independent experiments performed in sextuplicate. (C) Representative bright-field images of FaDu and Detroit-562 cells after 24-hour treatment with 2-APB at corresponding IC₅₀ concentrations. Representative images are shown to illustrate qualitative morphological changes (cell shrinkage, rounding, detachment) associated with cytotoxicity. Quantitative assessment of viability at 48 hours is provided in panels A-B. Images were captured at 10X magnification; scale bar = 500 µm.

Article Snippet: The HNSCC cell lines FaDu and Detroit-562 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: MTT Assay, Control

Journal: PLOS One

Article Title: Context-dependent dysregulation of store-operated calcium channels in head and neck squamous cell carcinoma

doi: 10.1371/journal.pone.0344393

Figure Lengend Snippet: (A, B) The effect of 2-APB on cell migration was assessed using a wound healing assay in FaDu (A) and Detroit-562 (B) cells. Cells were treated with 200 µM (FaDu) or 100 µM (Detroit-562) 2-APB for 12 hours. Representative bright-field images were captured at 0 and 12 hours using a light microscope at 5 × magnification. Scale bar = 200 µm. Migration rates (%) were calculated using ImageJ software with the formula: Migration (%) = [(Distance at 0 h – Distance at 12 h) / Distance at 0 h] × 100. Data represent the mean ± SEM of at least three independent experiments performed in triplicate. Statistical significance was assessed using Student’s t-test. *** p < 0.001.

Article Snippet: The HNSCC cell lines FaDu and Detroit-562 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Migration, Wound Healing Assay, Light Microscopy, Software

Journal: PLOS One

Article Title: Context-dependent dysregulation of store-operated calcium channels in head and neck squamous cell carcinoma

doi: 10.1371/journal.pone.0344393

Figure Lengend Snippet: (A) FaDu and Detroit-562 cells were exposed to continuous 2-APB treatment (200 µM and 100 µM, respectively) for 3–4 weeks to assess long-term clonogenic survival. Representative images show the absence of colony formation in treated cells compared to robust colony growth in untreated controls. (B) Cells were treated with 2-APB for 72 hours, after which the drug was removed and replaced with drug-free medium. Following a 3–4 weeks recovery period, colonies were fixed and stained. Colony area (%) was quantified using ImageJ, and results represent the mean ± SEM of at least three independent experiments. Statistical significance was assessed using a Student’s t-test. **p < 0.01.

Article Snippet: The HNSCC cell lines FaDu and Detroit-562 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Staining