recombinant human sclerostin sost  (R&D Systems)

Journal: Cells

Article Title: Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion

doi: 10.3390/cells13020137

Figure Lengend Snippet: Primer sequences.

Article Snippet: The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested.

Techniques: Sequencing

Journal: Cells

Article Title: Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion

doi: 10.3390/cells13020137

Figure Lengend Snippet: Clinical patient characteristics and H-score values derived from semiquantitative immunohistochemical evaluation of sclerostin expression at the OSCC–jawbone interface. pT indicates the extent of primary OSCC (pT2: tumor extent ≤2 cm with depth of invasion >5 mm and ≤10 mm; or tumor extent >2 cm and ≤4 cm with depth of invasion ≤10 mm; pT3: tumor extent >2 cm and ≤4 cm with depth of invasion >10 mm; or tumor extension >4 cm with depth of invasion ≤10 mm; pT4a: tumor extension >4 cm with depth of invasion >10 mm; or tumor invades adjacent structures (e.g., through the cortical bone of the mandible or maxilla). pN indicates cervical lymph node metastasis (pN0: no lymph node metastasis; pN2b: metastasis in multiple ipsilateral nodes, none >6 cm in largest dimension, and no extranodal extension. pN2c: metastasis in bilateral or contralateral lymph nodes, none >6 cm in largest dimension, and no extranodal extension; pN3b: metastasis in one or more nodes and clinically evident extranodal extension). pM indicates distant metastases (pM0: no distant metastases). H-score values range from 0 to 300, with 0 indicating that no cells are positive and 300 indicating that all cells are strongly positive.

Article Snippet: The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested.

Techniques: Derivative Assay, Immunohistochemistry, Expressing

Journal: Cells

Article Title: Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion

doi: 10.3390/cells13020137

Figure Lengend Snippet: Immunohistochemical staining protocol.

Article Snippet: The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested.

Techniques: Immunohistochemistry, Staining

Journal: Cells

Article Title: Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion

doi: 10.3390/cells13020137

Figure Lengend Snippet: Graphical representation of the averaged delta CT values from the qRT-PCR experiments for the two different OSCC tumor cell lines (SCC-040 and PCI-13). Higher CT values indicate low sclerostin expression, while lower values indicate higher expression. For normalization, the delta CT value was calculated with the CT values of GAPDH and sclerostin (SOST) from each cell line. SCC-040 tumor cells have higher sclerostin expression than PCI-13 tumor cells.

Article Snippet: The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested.

Techniques: Quantitative RT-PCR, Expressing

Journal: Cells

Article Title: Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion

doi: 10.3390/cells13020137

Figure Lengend Snippet: Graphical representation of the relative mRNA expression from the qRT-PCR experiments for the different osteogenesis markers ALPL, COL1A1, osteocalcin, osteopontin, osterix, and Runx2, as well as sclerostin expression. Undifferentiated hMSCs served as controls, GAPDH was used as internal housekeeping gene. Successful osteogenic differentiation of hMSCs is shown.

Article Snippet: The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested.

Techniques: Expressing, Quantitative RT-PCR

Journal: Cells

Article Title: Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion

doi: 10.3390/cells13020137

Figure Lengend Snippet: Illustration of tumor cell proliferation under sclerostin treatment at different concentrations (1, 5, and 10 ng/mL) and time periods (24 h, 36 h, 48 h, 60 h, and 72 h). Untreated tumor cells (0 ng/mL) were used as controls: ( a ) SCC-040 cell line; ( b ) PCI-13 cell line. Statistical test: one-way ANOVA with Tukey’s tests for post hoc comparisons; * p -values between 0.01 and 0.05 (significant); ** p -values between 0.001 and 0.01 (very significant). A significant influence of sclerostin on the tumor cell proliferation rate is shown, which seems to be time- and dose-dependent.

Article Snippet: The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested.

Techniques:

Journal: Cells

Article Title: Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion

doi: 10.3390/cells13020137

Figure Lengend Snippet: Illustration of PCI-13 tumor cell migration and invasion after 24 h under sclerostin treatment at different concentrations (1 and 5 ng/mL). Untreated tumor cells (0 ng/mL) were used as controls: ( a ) Migration of PCI-13 tumor cells; ( b ) Invasion of PCI-13 tumor cells. Statistical test: one-way ANOVA with Tukey’s tests for post hoc comparisons; ** p -values between 0.001 and 0.01 (very significant); **** p -values < 0.0001 (extremely significant). A significant effect of sclerostin on tumor cell migration and invasion rate is shown, which seems to be dose-dependent.

Article Snippet: The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested.

Techniques: Migration

Journal: Cells

Article Title: Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion

doi: 10.3390/cells13020137

Figure Lengend Snippet: Illustration of the scratch assay performed to verify migration characteristics for the PCI-13 OSCC tumor cell line with (1 and 5 ng/mL) and without (0 ng/mL, control) sclerostin treatment. Magnification ×4.

Article Snippet: The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested.

Techniques: Wound Healing Assay, Migration

Journal: Cells

Article Title: Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion

doi: 10.3390/cells13020137

Figure Lengend Snippet: Illustration of the scratch assay to visualize the migration rate of PCI-13 tumor cells under sclerostin treatment at different concentrations (1 and 5 ng/mL) over different time periods (0 h, 12 h, 24 h, 36 h). No significant effect of sclerostin was observed in the scratch assay.

Article Snippet: The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested.

Techniques: Wound Healing Assay, Migration

Journal: Cells

Article Title: Sclerostin Alters Tumor Cell Characteristics of Oral Squamous Cell Carcinoma and May Be a Key Player in Local Bone Invasion

doi: 10.3390/cells13020137

Figure Lengend Snippet: Histological illustration of the OSCC–jawbone interface with additional sclerostin immunohistochemistry. ( A ) Overview of the boundary between the local jawbone and the surrounding tumor cell clusters of OSCC; ( B ) enlarged view; ( C ) highly magnified image of direct tumor cell–bone contact with osteolysis lacunae; ( D ) visualization of multinucleated osteoclasts within osteolysis lacunae (OL). Significant sclerostin expression was found in both OSCC tumor cells and bone-associated cells at the OSCC–jawbone interface.

Article Snippet: The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested.

Techniques: Immunohistochemistry, Expressing