Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the Formyl Peptide Receptor type 1 to prevent the adhesion of ovarian cancer cells onto mesothelium and subsequent invasion

doi: 10.1186/s13046-019-1465-8

Figure Lengend Snippet: Clinicopathologic findings, uPAR and FPR1 expression in EOC tissues

Article Snippet: Cells (0.5 × 10 6 cells/sample) were detached using 200 mg/L EDTA, 500 mg/L trypsin (Cambrex) and incubated with 100 μl phosphate buffered saline (PBS) containing diluents (CTRL), 10 μl uPAR (CD87)-APC-conjugated antibody ( Miltenyi Biotec), or 30 μl FPR1 PE-conjugated Antibody ( R&D Sistems) for 10 min at 4 °C and 30 min at 23 °C, respectively.

Techniques: Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the Formyl Peptide Receptor type 1 to prevent the adhesion of ovarian cancer cells onto mesothelium and subsequent invasion

doi: 10.1186/s13046-019-1465-8

Figure Lengend Snippet: FPR1 requirement for adhesion of SKOV-3 cells onto vitronectin. a-b. Cell adhesion of SKOV-3 cells pre-incubated with serum-free medium (none), 100 nM SRSRY ( a ) or 100 nM fMLF ( b ) for 37 min at 37 °C in humidified air with 5%CO 2, onto the indicated matrix proteins. Cell adhesion was expressed as percentage of CTRL (cells adherent to plated coated with 1% BSA). Data are the means ± SD of three independent experiments with * P < 0.001. c. Adhesion onto Vn of SKOV-3 cells desensitized with diluents (none), 100 nM SRSRY, 100 nM fMLF, or 100 nM ARARY, monitored in real time for 2 h by the xCELLigence system. Data represent mean ± SD from a quadruplicate experiment representative of 3 replicates. d. Adhesion of SKOV-3 cells onto the indicated matrix proteins in the absence or presence of 10 nM RI-3. The number of adherent cells was expressed as a percentage of CTRL (cells adherent to plated coated with 1% BSA). Data are the means ± SD of three independent experiments with * P < 0.001. e. Adhesion of SKOV-3 cells onto Vn in the absence or presence of 10 nM RI-3, monitored in real time for 2 h by the xCELLigence system. Data represent mean ± SD from a quadruplicate experiment representative of 3 replicates

Article Snippet: Cells (0.5 × 10 6 cells/sample) were detached using 200 mg/L EDTA, 500 mg/L trypsin (Cambrex) and incubated with 100 μl phosphate buffered saline (PBS) containing diluents (CTRL), 10 μl uPAR (CD87)-APC-conjugated antibody ( Miltenyi Biotec), or 30 μl FPR1 PE-conjugated Antibody ( R&D Sistems) for 10 min at 4 °C and 30 min at 23 °C, respectively.

Techniques: Incubation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the Formyl Peptide Receptor type 1 to prevent the adhesion of ovarian cancer cells onto mesothelium and subsequent invasion

doi: 10.1186/s13046-019-1465-8

Figure Lengend Snippet: uPAR and FPR1 co-ordinated expression in human EOC tissues. Expression of uPAR and FPR1 in 5 HGSC tissues. Representative images of formalin-fixed, paraffin embedded sections subjected to immunohistochemical staining with R4 anti-uPAR mAb or anti-FPR1 Ab. Nuclei were stained blue with hematoxylin. Original magnification: × 200

Article Snippet: Cells (0.5 × 10 6 cells/sample) were detached using 200 mg/L EDTA, 500 mg/L trypsin (Cambrex) and incubated with 100 μl phosphate buffered saline (PBS) containing diluents (CTRL), 10 μl uPAR (CD87)-APC-conjugated antibody ( Miltenyi Biotec), or 30 μl FPR1 PE-conjugated Antibody ( R&D Sistems) for 10 min at 4 °C and 30 min at 23 °C, respectively.

Techniques: Expressing, Formalin-fixed Paraffin-Embedded, Immunohistochemical staining, Staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the Formyl Peptide Receptor type 1 to prevent the adhesion of ovarian cancer cells onto mesothelium and subsequent invasion

doi: 10.1186/s13046-019-1465-8

Figure Lengend Snippet: Generation of a primary cell culture from a human high-grade serous ovarian cancer . a HGSG tissue (patient # 5) processed for IHC analysis of uPAR and FPR1 expression on paraffin sections. Nuclei were stained blue with hematoxylin. Original magnifications: × 100. b Primary EOC cells derived from the same HGSG tissue visualized by phase contrast microscopy. Original magnification: × 400. c Western blotting of whole cell lysate from EOC cells with R4 anti-uPAR mAb, anti-FPR1 Ab, and anti-GAPDH Ab. d Bar graphs showing the average quantification of the uPAR/GAPDH and FPR1/GAPDH content from 3 independent experiments. Statistical significance with * P < 0.001. e Images of human EOC cells immunostained with R4 anti-uPAR mAb or anti-FPR1 Ab and visualized by a fluorescence inverted microscope. Nuclei were stained blue with DAPI. Original magnification: 1000x. f Fluorescence associated to EOC cells pre-incubated with diluents (none), 100 nM fMLF, 100 nM SRSRY or 100 nM RI-3, for 60 min at 4 °C, and then exposed to 10 nM FITC-RI3 for additional 60 min at 4 °C. Data represent mean +/− SD from three experiments performed in triplicate with * P < 0.001. g EOC cells exposed to 10 nM FITC-fMLF or 10 nM FITC-RI-3 for 30 min at 37 °C and then visualized using a Zeiss 510 Meta LSM microscope in 2D (left) or 3D (right) projections. Original magnification: 630×

Article Snippet: Cells (0.5 × 10 6 cells/sample) were detached using 200 mg/L EDTA, 500 mg/L trypsin (Cambrex) and incubated with 100 μl phosphate buffered saline (PBS) containing diluents (CTRL), 10 μl uPAR (CD87)-APC-conjugated antibody ( Miltenyi Biotec), or 30 μl FPR1 PE-conjugated Antibody ( R&D Sistems) for 10 min at 4 °C and 30 min at 23 °C, respectively.

Techniques: Cell Culture, Expressing, Staining, Derivative Assay, Microscopy, Western Blot, Fluorescence, Inverted Microscopy, Incubation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the Formyl Peptide Receptor type 1 to prevent the adhesion of ovarian cancer cells onto mesothelium and subsequent invasion

doi: 10.1186/s13046-019-1465-8

Figure Lengend Snippet: FPR1 expression in human EOC tissues. Ovarian cancer TMA slides were processed for IHC analysis of FPR1 expression. The three representative panels show intense ( a, d ), medium ( b, e ) and low ( c, f ) expression of FPR1 in primary ( a, b, c ) and metastatic ( d, e, f ) EOC tissues. Original magnifications: × 100 (left) and × 200 (right)

Article Snippet: Cells (0.5 × 10 6 cells/sample) were detached using 200 mg/L EDTA, 500 mg/L trypsin (Cambrex) and incubated with 100 μl phosphate buffered saline (PBS) containing diluents (CTRL), 10 μl uPAR (CD87)-APC-conjugated antibody ( Miltenyi Biotec), or 30 μl FPR1 PE-conjugated Antibody ( R&D Sistems) for 10 min at 4 °C and 30 min at 23 °C, respectively.

Techniques: Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the Formyl Peptide Receptor type 1 to prevent the adhesion of ovarian cancer cells onto mesothelium and subsequent invasion

doi: 10.1186/s13046-019-1465-8

Figure Lengend Snippet: Relationship between FPR1 expression and clinicopathological features of ovarian cancer tissues

Article Snippet: Cells (0.5 × 10 6 cells/sample) were detached using 200 mg/L EDTA, 500 mg/L trypsin (Cambrex) and incubated with 100 μl phosphate buffered saline (PBS) containing diluents (CTRL), 10 μl uPAR (CD87)-APC-conjugated antibody ( Miltenyi Biotec), or 30 μl FPR1 PE-conjugated Antibody ( R&D Sistems) for 10 min at 4 °C and 30 min at 23 °C, respectively.

Techniques: Expressing, Staining, Significance Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the Formyl Peptide Receptor type 1 to prevent the adhesion of ovarian cancer cells onto mesothelium and subsequent invasion

doi: 10.1186/s13046-019-1465-8

Figure Lengend Snippet: FPR1 expression in primary and metastatic ovarian cancer tissues

Article Snippet: Cells (0.5 × 10 6 cells/sample) were detached using 200 mg/L EDTA, 500 mg/L trypsin (Cambrex) and incubated with 100 μl phosphate buffered saline (PBS) containing diluents (CTRL), 10 μl uPAR (CD87)-APC-conjugated antibody ( Miltenyi Biotec), or 30 μl FPR1 PE-conjugated Antibody ( R&D Sistems) for 10 min at 4 °C and 30 min at 23 °C, respectively.

Techniques: Expressing, Staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the Formyl Peptide Receptor type 1 to prevent the adhesion of ovarian cancer cells onto mesothelium and subsequent invasion

doi: 10.1186/s13046-019-1465-8

Figure Lengend Snippet: Schematic diagram of proposed mechanism of RI-3 inhibitory effects on EOC cell adhesion. After detachment from the primary tumor, EOC cells float in the peritoneal fluid. EOC cells able to adhere to mesothelial ECM components via integrins may penetrate mesothelial cell layers. Intact uPAR or uPAR-derived fragments containing the 84–95 sequence that are expressed on EOC cell surface or may be released in the ascitic fluid, bind to FPR1 generating a cascade of events. Once engaged, FPR1 internalizes, triggering αvβ3 vitronectin receptor activation which, in turn, allows EOC cells to adhere onto mesothelial Vn. Subsequently, proteases within the mesothelial milieu allow EOC cells to invade the mesothelial cell layer where they proliferate and form metastases. This process may be counteracted by nanomolar concentrations of RI-3 peptide, blocking the assembly of uPAR/FPR1 complexes

Article Snippet: Cells (0.5 × 10 6 cells/sample) were detached using 200 mg/L EDTA, 500 mg/L trypsin (Cambrex) and incubated with 100 μl phosphate buffered saline (PBS) containing diluents (CTRL), 10 μl uPAR (CD87)-APC-conjugated antibody ( Miltenyi Biotec), or 30 μl FPR1 PE-conjugated Antibody ( R&D Sistems) for 10 min at 4 °C and 30 min at 23 °C, respectively.

Techniques: Derivative Assay, Sequencing, Activation Assay, Blocking Assay

Journal: Journal of Translational Medicine

Article Title: Defective formyl peptide receptor 2/3 and annexin A1 expressions associated with M2a polarization of blood immune cells in patients with chronic obstructive pulmonary disease

doi: 10.1186/s12967-018-1435-5

Figure Lengend Snippet: Changes in FPR1/2/3 expressions of blood immune cells in COPD patients after 1-year treatment and with various clinical phenotypes. In 10 COPD patients after 1-year treatment, a FPR3 expression of M1 monocyte, b NK cell, and c Th cell, as well as d FPR2 expression on Th cell all showed significant elevation. e COPD patients with a high MMRC dyspnea scale had higher FPR1 expression on neutrophil, while f those with a history of frequent moderate exacerbation in the past 1 year had lower FPR2 expression on neutrophil. g FPR1 expression on neutrophil showed significant reduction after 1-year treatment. h Ex-smokers with COPD had higher FPR3 expression of NK T cell than current smokers with COPD after 1 year treatment. COPD patient receiving oral steroid treatment for more than 3 months had i higher FPR2 expression on Th cell and j higher FPR3 expression of M1 monocyte than those without oral steroid use. *p < 0.05 for comparisons between COPD patients with a specific phenotype and healthy non-smokers by ANOVA test. # p < 0.05 for comparison between COPD patients with and without s specific phenotype by ANOVA test. § p < 0.05 for comparisons between COPD patients with and without a specific management after 1-year follow-up

Article Snippet: (A) Surface markers were measured by simultaneously staining for 30 min at 4 °C with directly conjugated monoclonal antibodies (mAb) as follows: PE Mouse Anti-Human CD56 (BD Pharmingen, USA), PE-CyTM5 Mouse Anti-Human CD3 (BD Pharmingen), CD4-PC7 (Beckman Coulter; USA), CD8-PC7, CD16-PC7, CD14-PC7 (Beckman Coulter), PerCP-Cy5.5 Mouse Anti-Human CD209 (BD Pharmingen), Anti-human CFS-conjugated FPR1 (R&D Systems; USA), PE-conjugated anti-human FPR2 (R&D Systems) or isotype control mAb. (B) For intracellular staining, cells were stained with anti-CD8a-PE-Cyanine5 and incubated with Cytofix/Cytoperm TM for 20 min at 4 °C, and washed with Perm/Wash Buffer.

Techniques: Expressing, Comparison