adamts1  (R&D Systems)

Journal: Cell death & disease

Article Title: Cyclic increase in the ADAMTS1-L1CAM-EGFR axis promotes the EMT and cervical lymph node metastasis of oral squamous cell carcinoma.

doi: 10.1038/s41419-024-06452-9

Figure Lengend Snippet: Fig. 1 ADAMTS1 expression promotes invasion of oral squamous cell carcinoma (SCC; OSCC) cells and correlates with poor survival outcomes in patients with head and neck SCC (HNSCC). A ADAMTS1 transcripts in normal and HNSCC tissues were analyzed using data from TCGA. Statistical significance was analyzed by a Wilcoxon signed-rank test. B Kaplan–Meier analysis of overall survival (OS) and disease-specific survival (DSS) rates in patients with HNSCC presenting with high or low ADAMTS1 expression using data from TCGA. C Endogenous protein levels of ADAMTS1 in OSCC cell lines (HSC-3, HSC-3M, SCC9, and SAS) were detected by a Western blot analysis. D In vitro invasive abilities of OSCC cell lines were analyzed by a Matrigel-invasion assay. E Western blot analysis of ADAMTS1 expressions in HSC-3 (left) and HSC-3M (right) cells respectively expressing ADAMTS1-flag and ADAMTS1 shRNAs. F Invasive abilities of HSC-3 and HSC-3M cells were determined by a Matrigel-invasion assay after infecting cells with a lentivirus carrying ADAMTS1-flag, ADAMTS1 shRNAs, ADAMTS1 shRNA co-expressing ADAMTS1-flag, or their respective control vectors. Upper panel: representative photomicrographs (200×). Lower panel: quantitative values from counting of invaded cells presented as the mean ± SD of three independent experiments. ***p < 0.001, compared to the respective control groups. ##p < 0.01, compared to ADAMTS1 shRNA-infected only group. G, H Proliferation rates and colony-formation capacities of ADAMTS1-manipulated HSC-3 and HSC-3M cells were respectively determined using MTS and colony-forming assays. ADAMTS1 expression had no obvious effect on cell proliferation rates during 24–96 h (G) or the colony-formation capacity during 7‒10 days (H). ns not significant.

Article Snippet: An antibody specific for ADAMTS1 (AF5867) was purchased from R&D Systems, while those for L1CAM (sc514360) and TGF-β (sc130348) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Western Blot, In Vitro, Invasion Assay, shRNA, Control, Infection

Journal: Cell death & disease

Article Title: Cyclic increase in the ADAMTS1-L1CAM-EGFR axis promotes the EMT and cervical lymph node metastasis of oral squamous cell carcinoma.

doi: 10.1038/s41419-024-06452-9

Figure Lengend Snippet: Fig. 2 ADAMTS1 expression accelerates tumor growth and cervical lymph node (LN) metastasis of oral squamous cell carcinoma (OSCC) in orthotopic mouse models. Male NOD/SCID mice were orthotopically injected with luciferase-tagged and ADAMTS1-overexpressing HSC-3 cells or ADAMTS1-knockdown HSC-3M cells. A, C Whole-body bioluminescence imaging was conducted each week for 5 weeks after injecting ADAMTS1-manipulated cells into mice. B, D Quantitative analysis of Xenogen imaging signal intensity (photons/s/cm2/sr) every week. *p < 0.05, ***p < 0.001, compared to the control group. Occurrence of cervical LN metastasis in NOD/SCID mice implanted with HSC-3/ADAMTS1 (E), HSC-3M/shADAMTS1 (F), or their respective control cells. The appearance, number and volume of cervical LNs were photographed, enumerated, and measured after removal. Data are presented as the mean ± SD. ***p < 0.001 compared to the control group.

Article Snippet: An antibody specific for ADAMTS1 (AF5867) was purchased from R&D Systems, while those for L1CAM (sc514360) and TGF-β (sc130348) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Injection, Luciferase, Knockdown, Imaging, Control

Journal: Cell death & disease

Article Title: Cyclic increase in the ADAMTS1-L1CAM-EGFR axis promotes the EMT and cervical lymph node metastasis of oral squamous cell carcinoma.

doi: 10.1038/s41419-024-06452-9

Figure Lengend Snippet: Fig. 3 L1CAM is critical in ADAMTS1-modulated invasive ability of oral squamous cell carcinoma (SCC; OSCC) cells and was correlated with poor prognoses in patients with head and neck SCC (HNSCC). A, B HSC-3 or HSC-3M cells expressed ADAMTS1-flag, shADAMTS1, or their respective control as indicated. Cell lysates and conditioned media were collected to detect endogenous ADAMTS1 or L1CAM protein levels (A) and secreted soluble L1CAM (B), respectively using western blotting and Dot-blotting assays. C A L1CAM shRNA was transfected into ADAMTS1-overexpressing HSC-3 cells as indicated and subjected to Matrigel-invasion assays. D HSC-3 cells were treated with indicated concentrations of rhL1CAM for 24 h and subjected to Matrigel-invasion assays. C, D Multiples of differences are presented as the mean ± SD of three independent experiments. ***p < 0.001, compared to the control group; ###p < 0.001, compared to the ADAMTS1-overexpressing only group. E L1CAM expression was analyzed in 43 matched HNSCC tissues and their corresponding normal tissues using data from TCGA. F Kaplan–Meier analysis of overall survival (OS) and disease-specific survival (DSS) rates in patients with HNSCC presenting with high or low L1CAM expression using data from TCGA. G Correlation analysis of TCGA HNSCC databases (TCGA, PanCancer Atlas) using cBioPortal which revealed a positive correlation between expressions of ADAMTS1 and L1CAM. H Survival heat map showing the prognostic impacts of ADAMTS1 and L1CAM on 33 different cancer types according to the GEPIA2 database. HR hazard ratio. I Combined high ADAMTS1 expression and high L1CAM expression were correlated with the worst OS and DSS in patients with HNSCC compared to patients with other expression statuses of ADAMTS1 and L1CAM.

Article Snippet: An antibody specific for ADAMTS1 (AF5867) was purchased from R&D Systems, while those for L1CAM (sc514360) and TGF-β (sc130348) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Control, Western Blot, shRNA, Transfection, Expressing

Journal: Cell death & disease

Article Title: Cyclic increase in the ADAMTS1-L1CAM-EGFR axis promotes the EMT and cervical lymph node metastasis of oral squamous cell carcinoma.

doi: 10.1038/s41419-024-06452-9

Figure Lengend Snippet: Fig. 6 Targeting ADAMTS1 by apigenin (API) resulted in suppression of the invasion of oral squamous cell carcinoma (OSCC) cells. A HSC-3, HSC-3M, and SAS cells were treated with API at 40 μM for different durations, and ADAMTS1, L1CAM, and EGFR expressions were evaluated by Western blotting. B HSC-3M cells were treated with API or infected with ADAMTS1 shRNAs for 24 h, and cell-invasive abilities were measured by a Matrigel-invasion assay. C, D HSC-3 cells were transiently transfected with a vector control or ADAMTS1-flag followed by API or vehicle treatment for an additional 24 h. ADAMTS1 expression and invasive ability in cells were respectively detected by Western blotting (C) and Matrigel-invasion assays (D). B, D Representative photographs of invaded cells (left panel) and quantification of those cells (right panel). Data are presented as the mean ± SD of three independent experiments, *p < 0.05, ***p < 0.001 vs. control cells and ##p < 0.01, vs. ADAMTS1-overexpressing only cells.

Article Snippet: An antibody specific for ADAMTS1 (AF5867) was purchased from R&D Systems, while those for L1CAM (sc514360) and TGF-β (sc130348) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Western Blot, Infection, Invasion Assay, Transfection, Plasmid Preparation, Control, Expressing

Journal: Cell death & disease

Article Title: Cyclic increase in the ADAMTS1-L1CAM-EGFR axis promotes the EMT and cervical lymph node metastasis of oral squamous cell carcinoma.

doi: 10.1038/s41419-024-06452-9

Figure Lengend Snippet: Fig. 8 Schematic presentation depicting the ADAMTS1-L1CAM-EGFR axis in promoting the epithelial-mesenchymal transition (EMT) and metastasis of oral squamous cell carcinoma (OSCC). EGFR activation might be triggered by formation of the ADAMTS1-L1CAM-EGFR complex or through ADAMTS1-mediated TGF-β upregulation to subsequently induce L1CAM upregulation and L1CAM-integrin binding, resulting in induction of IL-1β secretion. Bold dashed ovals indicate hypothetical molecules that participate in the ADAMTS1-L1CAM axis to transactivate EGFR signaling, and EGFR-activated signaling may exert positive feedback regulation on ADAMTS1 expression. Cyclic increases in ADAMTS1 and EGFR activation lead to exacerbation of the EMT and invasive abilities of OSCC cells.

Article Snippet: An antibody specific for ADAMTS1 (AF5867) was purchased from R&D Systems, while those for L1CAM (sc514360) and TGF-β (sc130348) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Activation Assay, Binding Assay, Expressing