oscc cell lines scc 9  (Procell Inc)

Journal: Chinese Medical Journal

Article Title: Precision therapy targeting CAMK2 to overcome resistance to EGFR inhibitors in FAT1-mutated oral squamous cell carcinoma

doi: 10.1097/cm9.0000000000003217

Figure Lengend Snippet: Figure 1: Mutational landscape of FAT1 in OSCC. Genetic mutation data of OSCC have been obtained from TCGA database (n = 328, https://portal.gdc.cancer.gov/). Oncoplot showed various information for each sample, including mutation types, mutation genes, and mutation burdens. The top of the figure showed the TMB of each sample. DEL: Dele tion; OSCC: Oral squamous cell carcinoma; TCGA: The Cancer Genome Atlas; TMB: Tumor mutational burden.

Article Snippet: The OSCC cell lines (SCC9, SCC25, SCC47, and HN12), which have been previously studied for their mutation characteristics,[24] were obtained from the American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Mutagenesis

Journal: Chinese Medical Journal

Article Title: Precision therapy targeting CAMK2 to overcome resistance to EGFR inhibitors in FAT1-mutated oral squamous cell carcinoma

doi: 10.1097/cm9.0000000000003217

Figure Lengend Snippet: Figure 2: KN93 enhances the therapeutic sensitivity of FAT1-mutated OSCC to afatinib. (A) The sensitivity of FAT1-mutated (SCC9 and SCC25) and FAT1-WT (HN12 and SCC47) OSCC cells to afatinib, and the sensitivity of FAT1-mutated cells to the combination of afatinib and KN93 (n = 4). (B) Representative fluorescence images and quantified results of EdU in FAT1-mutated cells (Scale bar = 100 µm). SCC9 and SCC25 cells were treated with 1 µmol/L afatinib, KN93 (30 µmol/L KN93 for SCC9, 10 µmol/L for SCC25), or their combination. (C) Early and late apoptosis in SCC9 and SCC25 cells after treating with 1 µmol/L afatinib, 30 µmol/L KN93, or their combination for 24 h. (D) Representative optical microscope images reflecting cell migration (Scale bar = 150 µm). SCC9 and SCC25 cells were treated with 1 µmol/L afatinib, 30 µmol/L KN93, or their combination for 6 h, and then migrated for 42 h under continuous drug exposure. (E, F) Representative images of xenograft tumors from SCC47 (E), SCC9 (F) cells isolated after single or combination treatment. All data were means ± SEM and were representative of ≥3 independent experiments (*P <0.05, †P <0.01, ‡P <0.001). CCK-8: Cell counting kit-8; CI: Confidence inteval; IC50: Half maximal inhibitory concentration; ns: Not significantly statistical difference; OSCC: Oral squamous cell carcinoma; SEM: Standard error of mean; WT: Wild type.

Article Snippet: The OSCC cell lines (SCC9, SCC25, SCC47, and HN12), which have been previously studied for their mutation characteristics,[24] were obtained from the American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Fluorescence, Microscopy, Migration, Isolation, CCK-8 Assay, Cell Counting, Concentration Assay

Journal: Chinese Medical Journal

Article Title: Precision therapy targeting CAMK2 to overcome resistance to EGFR inhibitors in FAT1-mutated oral squamous cell carcinoma

doi: 10.1097/cm9.0000000000003217

Figure Lengend Snippet: Figure 3: The combination of afatinib and KN93 inhibits the EGFR pathway and downregulates SOX2 and YAP1 in FAT1-mutated cells. (A,B) Western blot analysis of EGFR, pEGFR, MEK1/2, and pMEK expression in FAT1-mutated cells (SCC9, SCC25) (A) and FAT1-WT cells (SCC47, HN12) (B) after being treated with 10 µmol/L afatinib for 6 h. The EGF-treated groups were pre- treated with 10 ng/mL EGF for 30 min. (C, D) Western blot analysis of EGFR-RAS/RAF-MEK pathway protein expression in SCC25 and SCC9 cells after treatment with 10 µmol/L afatinib, 30 µmol/L KN93, and their combination for 6 h without (C) or with (D) pre-treatment with 10 ng/mL EGF for 30 min. (E) Representative IF images showing YAP1 location in FAT1-WT and mutated OSCC cells. (F) Expression of YAP1 and PYAP1 in FAT1-mutated and WT cells after being treated with 30 µmol/L KN93. (G) Representative IF images showing YAP1 translocation in FAT1-WT and mutated OSCC cells after being treated with 30 µmol/L KN93. (H) The mean optical density was quantified by ImageJ software from (G). (I) Western blot analysis of SOX2 after cells being treated with 30 µmol/L KN93 for 12 h. Scale bar = 100 µm. Data were mean ± SEM and were representative of ≥3 independent experiments. (*P <0.05, †P <0.01, ‡P <0.001). EGF: Epidermal growth factor; EGFR: Epidermal growth factor receptor; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; IF: Immunofluorescence; MEK: Mitogen-acti vated protein kinase kinase; MEK1/2: Mitogen-activated protein kinase kinase 1/2; NC: Negative control; ns: Not significantly statistical difference; OSCC; Oral squamous cell carcinoma; pEGFR: Phosphorylated epidermal growth factor receptor; pMEK: Phosphorylated mitogen-activated protein kinase kinase; SEM: Standard error of mean; SOX2: SRY-box transcription factor 2; WT: Wild-type; YAP1: Yes-associated protein 1.

Article Snippet: The OSCC cell lines (SCC9, SCC25, SCC47, and HN12), which have been previously studied for their mutation characteristics,[24] were obtained from the American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Western Blot, Expressing, Translocation Assay, Software, Immunofluorescence, Negative Control

Journal: Chinese Medical Journal

Article Title: Precision therapy targeting CAMK2 to overcome resistance to EGFR inhibitors in FAT1-mutated oral squamous cell carcinoma

doi: 10.1097/cm9.0000000000003217

Figure Lengend Snippet: Figure 6: The mechanism of combination therapy. In OSCC, FAT1 mutation can activate CAMK2, leading to the resistance to EGFR inhibitors (left). KN93 inhibited the activation of CAMK2, which blocked the suppression of EGFR signaling pathway, and also reduced the activity of YAP1 and SOX2. Moreover, the enhanced sensitivity of the EGFR-RAS/RAF- MEK pathway to afatinib enabled afatinib to elicit its anticancer effect. The combination treatment also induces mitochondria dysfunction and finally leads to FAT1 mutation OSCC cell death. CAMK2: Ca2+/calmodulin-dependent protein kinase II; EGFR: Epidermal growth factor receptor; FAT1: FAT atypical cadherin 1; MEK1/2: Mitogen-activated protein kinase kinase 1/2; OSCC: Oral squamous cell carcinoma; ROS: Reactive oxygen species; SOX2: SRY-box transcription factor 2; YAP1: Yes-associated protein 1.

Article Snippet: The OSCC cell lines (SCC9, SCC25, SCC47, and HN12), which have been previously studied for their mutation characteristics,[24] were obtained from the American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Mutagenesis, Activation Assay, Activity Assay