human fascin1  (Addgene inc)

Journal: International Journal of Oncology

Article Title: Fascin1 expression in high-grade serous ovarian carcinoma is a prognostic marker and knockdown of fascin1 suppresses the proliferation of ovarian cancer cells

doi: 10.3892/ijo.2013.2232

Figure Lengend Snippet: The association between fascin1 expression and clinicopathologic parameters of patients with high-grade ovarian serous carcinoma.

Article Snippet: After three rinses in phosphate-buffered saline (PBS) for 5 min, each section was incubated for 1 h at room temperature with mouse monoclonal antibodies to human fascin1 (Epitomics, Burlingame, CA, USA; 1:100), and human β-catenin (Novus Biologicals, Littleton, CO, USA) all diluted in PBS; washed three times in PBS for 5 min; incubated with horseradish peroxidase-labeled rabbit anti-mouse immunoglobulin (DAKO; 1 h at room temperature); washed three times; and incubated with a solution of diaminobenzidine (DAB) at room temperature to visualize peroxidase activity.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: Fascin1 expression in high-grade serous ovarian carcinoma is a prognostic marker and knockdown of fascin1 suppresses the proliferation of ovarian cancer cells

doi: 10.3892/ijo.2013.2232

Figure Lengend Snippet: Representative immunohistochemical analysis of the fascin1 and β-catenin expression. (A) Fascin1 was negative in the epithelial cells of the fallopian tube (negative control) and immunopositive in the endothelial cells (positive control) and (B) positive staining in cytoplasm of tumor cells of the high-grade ovarian serous carcinoma. β-catenin was observed clear membrane staining in all the cases of (C) fallopian tube and of (D) high-grade ovarian serous carcinoma.

Article Snippet: After three rinses in phosphate-buffered saline (PBS) for 5 min, each section was incubated for 1 h at room temperature with mouse monoclonal antibodies to human fascin1 (Epitomics, Burlingame, CA, USA; 1:100), and human β-catenin (Novus Biologicals, Littleton, CO, USA) all diluted in PBS; washed three times in PBS for 5 min; incubated with horseradish peroxidase-labeled rabbit anti-mouse immunoglobulin (DAKO; 1 h at room temperature); washed three times; and incubated with a solution of diaminobenzidine (DAB) at room temperature to visualize peroxidase activity.

Techniques: Immunohistochemical staining, Expressing, Negative Control, Positive Control, Staining, Membrane

Journal: International Journal of Oncology

Article Title: Fascin1 expression in high-grade serous ovarian carcinoma is a prognostic marker and knockdown of fascin1 suppresses the proliferation of ovarian cancer cells

doi: 10.3892/ijo.2013.2232

Figure Lengend Snippet: Kaplan-Meier survival analysis of progression-free survival in all patients according to fascin1 expression. Significant differences among the subgroups with positive (dimed line) and negative (bold line) fascin1 expression indicate poor outcomes in patients with fascin1 expression group. Fascin1 expression group was significantly correlated with shorter progression-free survival (P<0.001). The log-rank test yielded significant P-values.

Article Snippet: After three rinses in phosphate-buffered saline (PBS) for 5 min, each section was incubated for 1 h at room temperature with mouse monoclonal antibodies to human fascin1 (Epitomics, Burlingame, CA, USA; 1:100), and human β-catenin (Novus Biologicals, Littleton, CO, USA) all diluted in PBS; washed three times in PBS for 5 min; incubated with horseradish peroxidase-labeled rabbit anti-mouse immunoglobulin (DAKO; 1 h at room temperature); washed three times; and incubated with a solution of diaminobenzidine (DAB) at room temperature to visualize peroxidase activity.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: Fascin1 expression in high-grade serous ovarian carcinoma is a prognostic marker and knockdown of fascin1 suppresses the proliferation of ovarian cancer cells

doi: 10.3892/ijo.2013.2232

Figure Lengend Snippet: Fascin1 expression after siRNA transfection. (A) Real-time PCR analysis revealed the fascin1 expression was inhibited in the cancer cells transfected with fascin1 siRNA compared with control cancer cells (P<0.001). (B) Western blot analysis showed fascin1 siRNA inhibited fascin1 expression compared with control cancer cells. β-actin was detected as a loading control (P<0.001).

Article Snippet: After three rinses in phosphate-buffered saline (PBS) for 5 min, each section was incubated for 1 h at room temperature with mouse monoclonal antibodies to human fascin1 (Epitomics, Burlingame, CA, USA; 1:100), and human β-catenin (Novus Biologicals, Littleton, CO, USA) all diluted in PBS; washed three times in PBS for 5 min; incubated with horseradish peroxidase-labeled rabbit anti-mouse immunoglobulin (DAKO; 1 h at room temperature); washed three times; and incubated with a solution of diaminobenzidine (DAB) at room temperature to visualize peroxidase activity.

Techniques: Expressing, Transfection, Real-time Polymerase Chain Reaction, Control, Western Blot

Journal: International Journal of Oncology

Article Title: Fascin1 expression in high-grade serous ovarian carcinoma is a prognostic marker and knockdown of fascin1 suppresses the proliferation of ovarian cancer cells

doi: 10.3892/ijo.2013.2232

Figure Lengend Snippet: The inactivation of fascin1 inhibits proliferation and invasive ability of ovarian cancer cell lines SKOV3, and OVCAR3. (A) Cell proliferation was measured by colony forming assay after fascin1 siRNA transfection. The colony formation of the transfected cells grew significantly slower than control cells at 72 h. Colony numbers of transfected cells decreased significantly to 95.7% (SKOV3), 78.1% (OVCAR3) compared with that in control cells (72 h) (P<0.05). (B) Wound-healing assays were performed to examine the effect of fascin1 inactivation on cell migration (×100). The migration activities of the transfected cells were decreased 51.3% (SKOV3), and 55.3% (OVCAR3) compared to control cells at 16 h (P<0.05). (C) Matrigel invasion assays were performed to examine the effect of fascin1 on cell invasion (×100). Diagram of the cell count in the membrane. Fascin1 siRNA transfected cells led to 35.8% (SKOV3), and 31.1% (OVCAR3) decrease in the number of invasive cells (P<0.05). The above results are from 3 independent experiments.

Article Snippet: After three rinses in phosphate-buffered saline (PBS) for 5 min, each section was incubated for 1 h at room temperature with mouse monoclonal antibodies to human fascin1 (Epitomics, Burlingame, CA, USA; 1:100), and human β-catenin (Novus Biologicals, Littleton, CO, USA) all diluted in PBS; washed three times in PBS for 5 min; incubated with horseradish peroxidase-labeled rabbit anti-mouse immunoglobulin (DAKO; 1 h at room temperature); washed three times; and incubated with a solution of diaminobenzidine (DAB) at room temperature to visualize peroxidase activity.

Techniques: Transfection, Control, Migration, Cell Counting, Membrane