Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 was localized at the outer mitochondrial membrane in HNSCC cells. ( A-D ) HNSCC cells were subjected to immunofluorescent staining with MTUS1/Flag (green) and MitoTracker red staining (A) or COX IV (red) (B) or MFN1/2 (red) (C) or TOMM20 (red) (D) and counterstained with DAPI (blue). ( E ) Equal amounts of whole cell lysate, mitochondria and cytoplasmic, membrane, soluble nuclear and chromatin-bound extractions from HNSCC cells were analyzed by western blotting using the indicated antibodies including mitochondria marker (COX IV and Timm23), MOM marker (MFN2 and TOMM20) and Golgi marker (GM130). ( F ) Mitochondria were isolated and incubated with protease K for 5 min and then subjected to western blotting to detect ATIP1, mitochondria marker (COX IV and Timm23) and MOM marker (MFN2 and TOMM20). ( G ) Cell lysates from HNSCC cells transduced with lentivirus containing ATIP1-Flag were subjected to immunoprecipitation with an antibody against Flag, followed by western blotting with the indicated antibodies (MFN2 or TOMM20). ( H ) Cell lysates were subjected to western blotting with the MFN2 or TOMM20 antibodies in MTUS1/ATIP1-overexpressed or control HNSCC cells. Scale bar, 10 mm. β-actin or GAPDH was used as loading proteins.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Membrane, Staining, Western Blot, Marker, Isolation, Incubation, Transduction, Immunoprecipitation, Control

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 influenced mitochondrial function and metabolism in HNSCC cells. ( A-B ) MitoTracker Red staining (A) or immunofluorescence staining with COX IV (B) were used to detect the distribution of mitochondria under a confocal microscope in MTUS1/ATIP1-overexpressed or control HNSCC cells. Scale bars, 20 μm. ( C ) Mitochondrial real-time motions of HNSCC cells, stained by MitoTracker Red were detected under a confocal time-lapse live-cell analysis system. White arrowheads indicate direction of movement of single moving mitochondria. ( D-E ) MTUS1/ATIP1-overexpressed or control SCC9 cells was determined using oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) assays. ( F ) Intracellular ATP levels were measured by a bioluminescence ATP determination assay in MTUS1/ATIP1-overexpressed or control HNSCC cells. ( G ) MTUS1/ATIP1-overexpressed or control HNSCC cells were stained with DCFH-DA and then ROS levels were detected using flow cytometry. ( H ) The MMP was detected by a fluorescence microscope using JC-1 probe staining in MTUS1/ATIP1-overexpressed or control HNSCC cells. All data are presented as the mean ± SEM of three independent experiments. ns, not statistically significant ( P ≥ 0.05); *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Staining, Immunofluorescence, Microscopy, Control, Cell Analysis, Flow Cytometry, Fluorescence

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 overexpression induced mitochondrial pyroptosis in HNSCC cells. ( A-C ) MTUS1/ATIP1-overexpressed or -knockdown HNSCC SCC9 cells were detected by high-content Microscopy imaging, LDH release assay and pyroptotic markers assay. MTUS1/ATIP1 overexpression induced pyroptosis (A), LDH release (B) and GSDME cleavage and Cas-9/-3 cleavage (C). ( D-F ) MTUS1/ATIP1-overexpressed SCC9 cells were treated with Cas-9 inhibitor Z-LEVD-FMK (20 μM) or Cas-3 inhibitor Z-LEHD-FMK (20 μM) for 2 h. Z-LEHD-FMK could lead to the blockade of Cas-9 and -3 cleavage. Z-DEVD-FMK could only lead to the blockade of Cas-3 cleavage. Cas-9/-3 inhibitors blocked MTUS1/ATIP1-induced GSDME cleavage (D), pyroptosis (E) and LDH release (F). Arrowheads indicate the large bubbles emerging from the plasma membrane. Scale bar, 20 μm. GSDME, gasdermin E; GSDMD, gasdermin D; FL, full-length; -N, cleaved N-terminal; -C, cleaved C-terminal; Pro-Cas, pro-caspase; Cl, cleaved. β-actin or GAPDH was used as loading proteins. All data are presented as the mean ± SEM of three independent experiments. ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Over Expression, Knockdown, Microscopy, Imaging, Lactate Dehydrogenase Assay, Clinical Proteomics, Membrane

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 induced ROS-dependent mitochondrial pyroptosis in HNSCC cells. MTUS1/ATIP1-overexpressed HNSCC SCC9 cells treated with or without ROS inhibitors NAC (5 mM) for 2h were used to detect Cyto c release, Cas-9/-3 cleavage, pyroptotic features (including LDH release and GSDME cleavage) and ROS level. ( A-B ) MTUS1/ATIP1-induced Cyto c release from mitochondria to cytosol was abolished by NAC in SCC9 cells. Cyto c release was detected by confocal microscopy (A) or western blotting in the cytosol fraction (B). Scale bar, 20 μm. ( C-D ) Cleavage of Cas-9 and -3 (C) and LDH release (D) induced by MTUS1/ATIP1 overexpression can be abolished by NAC in SCC9 cells. ( E-F ) Cas-9 and -3 inhibitor had no effect on the MTUS1/ATIP1-overexpressed-induced ROS elevation (E) and Cyto c release (F) in SCC9 cells. Cyto c, cytochrome c; Cl, cleaved; GSDME, gasdermin E; FL, full-length; -N, cleaved N-terminal. β-actin or GAPDH was used as loading proteins. All data are presented as the mean ± SEM of three independent experiments. ns, not statistically significant ( P ≥ 0.05), ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Confocal Microscopy, Western Blot, Over Expression

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 interacted with MFN2 and led to ROS-induced mitochondrial pyroptosis in HNSCC cells. ( A ) MFN2 knockdown blocked MTUS1/ATIP1 induced mitochondrial collapse. Scale bar, 20 μm. ( B-C ) MFN2 knockdown inhibited MTUS1/ATIP1-induced Cyto c release, Cas-9 and -3 cleavage (B) and ROS elevation (C) in HNSCC SCC9 cells. ( D-F ) MFN2 knockdown inhibited MTUS1/ATIP1-induced pyroptosis (D), GSDME cleavage (E) and LDH release (F) in HNSCC SCC9 cells. Scale bar, 100 μm. Cyto c, cytochrome c; Cl, cleaved; GSDME, gasdermin E; FL, full-length; -N, cleaved N-terminal. β-actin or GAPDH was used as loading proteins. All data are presented as the mean ± SEM of three independent experiments. ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Knockdown

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MFN2 recruited BAX to mitochondria and induced pyroptosis in MTUS1/ATIP1 overexpression HNSCC cells. ( A-B ) MFN2 knockdown blocked the translocation of BAX to mitochondria by confocal microscopy (A) and western blotting (B) in MTUS1/ATIP1 overexpression HNSCC cells. Scale bar, 10 μm. ( C-F ) BAX knockdown inhibited the MTUS1/ATIP1-induced mitochondrial collapse (C), Cyto c release, Cas-9 and Cas-3 cleavage, GSDME cleavage (D), pyroptosis (E) and the LDH release (F) in MTUS1/ATIP1 overexpression SCC9 cells. Scale bar, 100 μm (C). Scale bar, 5 μm (E). ( G ) BAX had no effect on the MTUS1/ATIP1-induced ROS elevation in MTUS1/ATIP1 overexpression SCC9 cells. Cyto c, cytochrome c; Cl, cleaved; GSDME, gasdermin E; FL, full-length; -N, cleaved N-terminal. β-actin or GAPDH was used as loading proteins. All data are presented as the mean ± SEM of three independent experiments. ns, not statistically significant ( P ≥ 0.05), ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Over Expression, Knockdown, Translocation Assay, Confocal Microscopy, Western Blot

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 exerted anticancer effects on HNSCC in vitro and in vivo . ( A ) MTUS1/ATIP1 overexpression inhibited the cell proliferation, migration and colony formation abilities in SCC9 cells. ( B ) MTUS1/ATIP1 knockdown enhanced the cell proliferation, migration and colony formation abilities in SCC9 cells. ( C and F ) Representative fluorescence intensity images of tumor-bearing mice, photograph of xenografts and its growth curves, and tumor weights in subcutaneous xenograft of MTUS1/ATIP1 overexpressed (C) or knockdowned mice (F). ( D and G ) Representative fluorescence intensity images of tumor-bearing mice, images of tongue lesions and H&E-staining image in in sito tongue tumor of MTUS1/ATIP1 overexpressed (D) or knockdowned mice (G) . Arrow indicates the location of palpable tongue tumor. ( E and H ) Representative the proportion of mice with lymph node metastases in lymph node metastasis model of MTUS1/ATIP1 overexpressed (E) or knockdowned mice (H). All data are presented as the mean ± SEM of three independent experiments. **P < 0.01; ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: In Vitro, In Vivo, Over Expression, Migration, Knockdown, Fluorescence, Staining

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 exerted antitumor effects in HNSCC PDX mouse models. ( A ) Schema for the establishment of HNSCC PDX models. Mice were treated by intratumoral injection of 10 8 TU of Lv-OE-ATIP1 or Lv-Ctrl once a week for 4 times. ( B-C ) Representative of xenografts photograph, growth curves and tumor weights in Lv-OE-ATIP1 treated HNSCC PDX models generated from two HNSCC patient samples (PDX HNSCC-1 and PDX HNSCC-2). ( D ) H&E staining showed a large area of tumour tissue necrosis (Arrow) in Lv-OE-ATIP1 treated group. ( E ) No significant differences in body weight was found between Lv-Ctrl and Lv-OE-ATIP1 groups. Data are presented as the mean ± SD (n = 7). *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Injection, Generated, Staining

Journal: American Journal of Translational Research

Article Title: Linc00467 promotes invasion and inhibits apoptosis of head and neck squamous cell carcinoma by regulating miR-1285-3p/TFAP2A

doi:

Figure Lengend Snippet: Expressions of Linc00467 and miR-1285-3p in HNSCC. A: Expressions of Linc00467 in the normal tissues adjacent to the tumor and cancer tissues; B: Expressions of miR-1285-3p in the normal tissues adjacent to the tumor and cancer tissues; C: Expressions of Linc00467 in the cell lines; D: Expressions of miR-1285-3p in the cell lines. Compared with the Normal group, △P<0.05; compared with HOK, #P<0.05; compared with SCC-9, @P<0.05.

Article Snippet: The HNSCC cell lines (HN4, HN6, SCC-4, SCC-9) and normal endocystal cell line HOK (ATCC cell bank, USA) were placed into the DMEM medium (including 10% fetal bovine serum) (PM150210, Shanghai Pure Biotech Co., Ltd., China).

Techniques:

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 was localized at the outer mitochondrial membrane in HNSCC cells. ( A-D ) HNSCC cells were subjected to immunofluorescent staining with MTUS1/Flag (green) and MitoTracker red staining (A) or COX IV (red) (B) or MFN1/2 (red) (C) or TOMM20 (red) (D) and counterstained with DAPI (blue). ( E ) Equal amounts of whole cell lysate, mitochondria and cytoplasmic, membrane, soluble nuclear and chromatin-bound extractions from HNSCC cells were analyzed by western blotting using the indicated antibodies including mitochondria marker (COX IV and Timm23), MOM marker (MFN2 and TOMM20) and Golgi marker (GM130). ( F ) Mitochondria were isolated and incubated with protease K for 5 min and then subjected to western blotting to detect ATIP1, mitochondria marker (COX IV and Timm23) and MOM marker (MFN2 and TOMM20). ( G ) Cell lysates from HNSCC cells transduced with lentivirus containing ATIP1-Flag were subjected to immunoprecipitation with an antibody against Flag, followed by western blotting with the indicated antibodies (MFN2 or TOMM20). ( H ) Cell lysates were subjected to western blotting with the MFN2 or TOMM20 antibodies in MTUS1/ATIP1-overexpressed or control HNSCC cells. Scale bar, 10 mm. β-actin or GAPDH was used as loading proteins.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Membrane, Staining, Western Blot, Marker, Isolation, Incubation, Transduction, Immunoprecipitation, Control

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 influenced mitochondrial function and metabolism in HNSCC cells. ( A-B ) MitoTracker Red staining (A) or immunofluorescence staining with COX IV (B) were used to detect the distribution of mitochondria under a confocal microscope in MTUS1/ATIP1-overexpressed or control HNSCC cells. Scale bars, 20 μm. ( C ) Mitochondrial real-time motions of HNSCC cells, stained by MitoTracker Red were detected under a confocal time-lapse live-cell analysis system. White arrowheads indicate direction of movement of single moving mitochondria. ( D-E ) MTUS1/ATIP1-overexpressed or control SCC9 cells was determined using oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) assays. ( F ) Intracellular ATP levels were measured by a bioluminescence ATP determination assay in MTUS1/ATIP1-overexpressed or control HNSCC cells. ( G ) MTUS1/ATIP1-overexpressed or control HNSCC cells were stained with DCFH-DA and then ROS levels were detected using flow cytometry. ( H ) The MMP was detected by a fluorescence microscope using JC-1 probe staining in MTUS1/ATIP1-overexpressed or control HNSCC cells. All data are presented as the mean ± SEM of three independent experiments. ns, not statistically significant ( P ≥ 0.05); *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Staining, Immunofluorescence, Microscopy, Control, Cell Analysis, Flow Cytometry, Fluorescence

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 overexpression induced mitochondrial pyroptosis in HNSCC cells. ( A-C ) MTUS1/ATIP1-overexpressed or -knockdown HNSCC SCC9 cells were detected by high-content Microscopy imaging, LDH release assay and pyroptotic markers assay. MTUS1/ATIP1 overexpression induced pyroptosis (A), LDH release (B) and GSDME cleavage and Cas-9/-3 cleavage (C). ( D-F ) MTUS1/ATIP1-overexpressed SCC9 cells were treated with Cas-9 inhibitor Z-LEVD-FMK (20 μM) or Cas-3 inhibitor Z-LEHD-FMK (20 μM) for 2 h. Z-LEHD-FMK could lead to the blockade of Cas-9 and -3 cleavage. Z-DEVD-FMK could only lead to the blockade of Cas-3 cleavage. Cas-9/-3 inhibitors blocked MTUS1/ATIP1-induced GSDME cleavage (D), pyroptosis (E) and LDH release (F). Arrowheads indicate the large bubbles emerging from the plasma membrane. Scale bar, 20 μm. GSDME, gasdermin E; GSDMD, gasdermin D; FL, full-length; -N, cleaved N-terminal; -C, cleaved C-terminal; Pro-Cas, pro-caspase; Cl, cleaved. β-actin or GAPDH was used as loading proteins. All data are presented as the mean ± SEM of three independent experiments. ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Over Expression, Knockdown, Microscopy, Imaging, Lactate Dehydrogenase Assay, Clinical Proteomics, Membrane

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 induced ROS-dependent mitochondrial pyroptosis in HNSCC cells. MTUS1/ATIP1-overexpressed HNSCC SCC9 cells treated with or without ROS inhibitors NAC (5 mM) for 2h were used to detect Cyto c release, Cas-9/-3 cleavage, pyroptotic features (including LDH release and GSDME cleavage) and ROS level. ( A-B ) MTUS1/ATIP1-induced Cyto c release from mitochondria to cytosol was abolished by NAC in SCC9 cells. Cyto c release was detected by confocal microscopy (A) or western blotting in the cytosol fraction (B). Scale bar, 20 μm. ( C-D ) Cleavage of Cas-9 and -3 (C) and LDH release (D) induced by MTUS1/ATIP1 overexpression can be abolished by NAC in SCC9 cells. ( E-F ) Cas-9 and -3 inhibitor had no effect on the MTUS1/ATIP1-overexpressed-induced ROS elevation (E) and Cyto c release (F) in SCC9 cells. Cyto c, cytochrome c; Cl, cleaved; GSDME, gasdermin E; FL, full-length; -N, cleaved N-terminal. β-actin or GAPDH was used as loading proteins. All data are presented as the mean ± SEM of three independent experiments. ns, not statistically significant ( P ≥ 0.05), ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Confocal Microscopy, Western Blot, Over Expression

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 interacted with MFN2 and led to ROS-induced mitochondrial pyroptosis in HNSCC cells. ( A ) MFN2 knockdown blocked MTUS1/ATIP1 induced mitochondrial collapse. Scale bar, 20 μm. ( B-C ) MFN2 knockdown inhibited MTUS1/ATIP1-induced Cyto c release, Cas-9 and -3 cleavage (B) and ROS elevation (C) in HNSCC SCC9 cells. ( D-F ) MFN2 knockdown inhibited MTUS1/ATIP1-induced pyroptosis (D), GSDME cleavage (E) and LDH release (F) in HNSCC SCC9 cells. Scale bar, 100 μm. Cyto c, cytochrome c; Cl, cleaved; GSDME, gasdermin E; FL, full-length; -N, cleaved N-terminal. β-actin or GAPDH was used as loading proteins. All data are presented as the mean ± SEM of three independent experiments. ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Knockdown

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MFN2 recruited BAX to mitochondria and induced pyroptosis in MTUS1/ATIP1 overexpression HNSCC cells. ( A-B ) MFN2 knockdown blocked the translocation of BAX to mitochondria by confocal microscopy (A) and western blotting (B) in MTUS1/ATIP1 overexpression HNSCC cells. Scale bar, 10 μm. ( C-F ) BAX knockdown inhibited the MTUS1/ATIP1-induced mitochondrial collapse (C), Cyto c release, Cas-9 and Cas-3 cleavage, GSDME cleavage (D), pyroptosis (E) and the LDH release (F) in MTUS1/ATIP1 overexpression SCC9 cells. Scale bar, 100 μm (C). Scale bar, 5 μm (E). ( G ) BAX had no effect on the MTUS1/ATIP1-induced ROS elevation in MTUS1/ATIP1 overexpression SCC9 cells. Cyto c, cytochrome c; Cl, cleaved; GSDME, gasdermin E; FL, full-length; -N, cleaved N-terminal. β-actin or GAPDH was used as loading proteins. All data are presented as the mean ± SEM of three independent experiments. ns, not statistically significant ( P ≥ 0.05), ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Over Expression, Knockdown, Translocation Assay, Confocal Microscopy, Western Blot

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 exerted anticancer effects on HNSCC in vitro and in vivo . ( A ) MTUS1/ATIP1 overexpression inhibited the cell proliferation, migration and colony formation abilities in SCC9 cells. ( B ) MTUS1/ATIP1 knockdown enhanced the cell proliferation, migration and colony formation abilities in SCC9 cells. ( C and F ) Representative fluorescence intensity images of tumor-bearing mice, photograph of xenografts and its growth curves, and tumor weights in subcutaneous xenograft of MTUS1/ATIP1 overexpressed (C) or knockdowned mice (F). ( D and G ) Representative fluorescence intensity images of tumor-bearing mice, images of tongue lesions and H&E-staining image in in sito tongue tumor of MTUS1/ATIP1 overexpressed (D) or knockdowned mice (G) . Arrow indicates the location of palpable tongue tumor. ( E and H ) Representative the proportion of mice with lymph node metastases in lymph node metastasis model of MTUS1/ATIP1 overexpressed (E) or knockdowned mice (H). All data are presented as the mean ± SEM of three independent experiments. **P < 0.01; ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: In Vitro, In Vivo, Over Expression, Migration, Knockdown, Fluorescence, Staining

Journal: International Journal of Biological Sciences

Article Title: Mitochondrial outer membrane protein MTUS1/ATIP1 exerts antitumor effects through ROS-induced mitochondrial pyroptosis in head and neck squamous cell carcinoma

doi: 10.7150/ijbs.94795

Figure Lengend Snippet: MTUS1/ATIP1 exerted antitumor effects in HNSCC PDX mouse models. ( A ) Schema for the establishment of HNSCC PDX models. Mice were treated by intratumoral injection of 10 8 TU of Lv-OE-ATIP1 or Lv-Ctrl once a week for 4 times. ( B-C ) Representative of xenografts photograph, growth curves and tumor weights in Lv-OE-ATIP1 treated HNSCC PDX models generated from two HNSCC patient samples (PDX HNSCC-1 and PDX HNSCC-2). ( D ) H&E staining showed a large area of tumour tissue necrosis (Arrow) in Lv-OE-ATIP1 treated group. ( E ) No significant differences in body weight was found between Lv-Ctrl and Lv-OE-ATIP1 groups. Data are presented as the mean ± SD (n = 7). *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The HNSCC cell lines SCC9, SCC15 and SCC25 were purchased from ATCC and HSC3 was purchased from the cell bank of the Japanese Collection of Research Bioresource (CRB), with the cell line STR authentication report.

Techniques: Injection, Generated, Staining

Journal: Cell Death & Disease

Article Title: LINC01123 promotes immune escape by sponging miR-214-3p to regulate B7–H3 in head and neck squamous-cell carcinoma

doi: 10.1038/s41419-022-04542-0

Figure Lengend Snippet: A Heat map with hierarchical clustering. Blue and red indicate low and high expression levels, respectively. B Venn diagram of upregulated LncRNAs in four HNSCC cell lines (CAL-27, Fadu, SCC-9, SCC-15) compared with HOEC. C TCGA data of LINC01123 expression in the classical subtype HNSCC tissues ( n = 49) and normal tissues ( n = 44). D The overall survival rate analysis of LINC01123. E Expression of LINC01123 in HNSCC cell lines (SCC-9, SCC-15, SCC-25, Fadu, CAL-27) and HOEC. F Expression of LINC01123 in HNSCC patient tissues and paracancerous tissues, as detected by qRT-PCR. G TCGA data of B7–H3 expression in HNSCC tissues ( n = 519) and normal tissues ( n = 44). H The overall survival rate analysis of B7–H3 . I B7–H3 is highly expressed in various types of tumor (COAD, DLBC, ESCA, GBM, KIRC, KIRP, LGG, LUSC, PAAD, SKCM, STAD, TGCT, THYM, and UCS). Each dot represents expression of samples. J Subcellular location of LINC01123, as detected by FISH assay. K Prediction of potential miRNA link between LINC01123 and B7–H3 . COAD colon adenocarcinoma, DLBC lymphoid neoplasm diffuse large b-cell lymphoma, ESCA esophageal carcinoma, GBM glioblastoma multiforme, KIRC kidney renal clear cell carcinoma, KIRP kidney renal papillary cell carcinoma, LGG brain lower grade glioma, LUSC lung squamous cell carcinoma, PAAD pancreatic adenocarcinoma, SKCM skin cutaneous melanoma, STAD stomach adenocarcinoma, TGCT testicular germ cell tumors, THYM thymoma, UCS uterine carcinosarcoma. The experiment was repeated three times.

Article Snippet: An HNSCC cell line (SCC-9, SCC-15) and 293 T cells were initially purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Current Oncology

Article Title: The Association of OLFM4 with the Progression and Cisplatin Resistance of Head and Neck Squamous Carcinoma

doi: 10.3390/curroncol32050276

Figure Lengend Snippet: OLFM4 promoted proliferation, invasion, migration, and cisplatin resistance in HNSCC. ( A ) Expression of OLFM4 in different HNSCC cell lines and a normal cell line; ( B ) qRT-PCR and WB evaluating lentiviral transfection efficiency in CAL27 and 5-8F cells with overexpressed or knocked-down OLFM4; ( C ) association of OLFM4 expression with proliferation ability of CAL27 and 5-8F cells (CCK8 assay); ( D ) association of OLFM4 expression with migration ability of CAL27 and 5-8F cells (scratch assay); ( E ) association of OLFM4 expression with invasion ability of CAL27 and 5-8F cells (transwell assay); ( F ) measurements of IC50 of cisplatin in CAL27 and 5-8F cells; ( G ) experimental steps for the co-culture of GFP-labeled 5-8F cells of each group (5-8F-OE-OLFM4, 5-8F-shOLFM4, 5-8F-OE-NC, 5-8F-shNC cells) with wild-type 5-8F cells (blank control without GFP-labeling), respectively; ( H ) fluorescence intensity detected by flow cytometry in 5-8F-OE-OLFM4 cells co-cultured with wild-type 5-8F cells and 5-8F-shOLFM4 cells co-cultured with wild-type 5-8F cells under cisplatin pressure at different concentrations. Notes: b, blank control without GFP labeling; OE, overexpression; NC, overexpression negative control; sh, knockdown; shNC, knockdown negative control; GFP, green fluorescent protein; * p < 0.05, ** p < 0.01, *** p < 0.001; **** p < 0.0001.

Article Snippet: Human HNSCC cell lines (5-8F, CAL27, SAS, SCC-9), normal oral keratinocytes (NOKs), and patient-derived models (HNSCC-PDOs, HNSCC-PDO cis-R ) were maintained in the Guangxi Key Laboratory for Early Prevention and Treatment of High-Incidence Tumors.

Techniques: Migration, Expressing, Quantitative RT-PCR, Transfection, CCK-8 Assay, Wound Healing Assay, Transwell Assay, Co-Culture Assay, Labeling, Control, Fluorescence, Flow Cytometry, Cell Culture, Over Expression, Negative Control, Knockdown

Journal: Oncology Research

Article Title: Fibroblast activation protein (FAP) as a prognostic biomarker in multiple tumors and its therapeutic potential in head and neck squamous cell carcinoma

doi: 10.32604/or.2024.046965

Figure Lengend Snippet: Overexpression of FAP was associated with poorer prognosis in HNSC patients. (a) The expression of FAP mRNA in 40 pairs of HNSC tissues and adjacent normal tissues. (b) Univariate analysis of FAP expression and also clinicopathological factors in TCGA patients with HNSC. (c) Multifactorial analysis of FAP expression and clinicopathological factors in TCGA patients with HNSC. (d) Nomogram for prognostic prediction. (e) The ROC curve of the nomogram predicting survival prognosis.

Article Snippet: The Human oral keratinocytes (HOK) and HNSC cell lines SCC9, SCC15, HN4, SCC25, and CAL27 were provided by the CCTCC.

Techniques: Over Expression, Expressing

Journal: Oncology Research

Article Title: Fibroblast activation protein (FAP) as a prognostic biomarker in multiple tumors and its therapeutic potential in head and neck squamous cell carcinoma

doi: 10.32604/or.2024.046965

Figure Lengend Snippet: FAP mediated the PI3K-Akt signal pathway in HNSC cells. (a) The expression of FAP mRNA in HOK cell line and HNSCs cell lines (SCC15, CAL27, SCC9, HN4, and SCC25). (b) Western blotting was used to detect the expression of FAP protein in HOK and HNSC cells. (c) Western blot analysis of the expression of FAP and PI3K-Akt-related molecules with FAP overexpression or knock-down in HNSC cells. *** p < 0.001.

Article Snippet: The Human oral keratinocytes (HOK) and HNSC cell lines SCC9, SCC15, HN4, SCC25, and CAL27 were provided by the CCTCC.

Techniques: Expressing, Western Blot, Over Expression, Knockdown

Journal: Oncology Research

Article Title: Fibroblast activation protein (FAP) as a prognostic biomarker in multiple tumors and its therapeutic potential in head and neck squamous cell carcinoma

doi: 10.32604/or.2024.046965

Figure Lengend Snippet: FAP overexpression promote the proliferation, migration and invasion of HNSC cells. (a) The results of real-time PCR showing FAP mRNA expression in HN4 cell after transfection of FAP knock-down vector for 48 h (*** p < 0.001). (b) FAP knockdown in HN4 cells inhibited the the proliferative ability of tumor cells (** p < 0.01). (c) FAP knockdown in HN4 cells inhibited the migration and invasion of tumor cells (*** p < 0.001). (d) FAP knockdown in HN4 cells inhibited the migration of tumor cells (*** p < 0.001).

Article Snippet: The Human oral keratinocytes (HOK) and HNSC cell lines SCC9, SCC15, HN4, SCC25, and CAL27 were provided by the CCTCC.

Techniques: Over Expression, Migration, Real-time Polymerase Chain Reaction, Expressing, Transfection, Knockdown, Plasmid Preparation

Journal: Oncology Research

Article Title: Fibroblast activation protein (FAP) as a prognostic biomarker in multiple tumors and its therapeutic potential in head and neck squamous cell carcinoma

doi: 10.32604/or.2024.046965

Figure Lengend Snippet: FAP knock-down inhibite the proliferation, migration and invasion of HNSC cells. (a) The expression of FAP mRNA in CAL27 cell after transfection of overexpression vector of FAP for 48 h (*** p < 0.001). (b) Overexpression of FAP in CAL27 cells facilitated the proliferation were detected in tumor cells. (** p < 0.01). (c) Overexpression of FAP in CAL27 cells promoted the migration and invasion of tumor cells (*** p < 0.001). (d) Overexpression of FAP in CAL27 cells facilitated the migration of tumor cells (*** p < 0.001).

Article Snippet: The Human oral keratinocytes (HOK) and HNSC cell lines SCC9, SCC15, HN4, SCC25, and CAL27 were provided by the CCTCC.

Techniques: Knockdown, Migration, Expressing, Transfection, Over Expression, Plasmid Preparation