Journal: Cell Transplantation

Article Title: Cyclooxygenase 2 Promotes Proliferation and Invasion in Ovarian Cancer Cells via the PGE2/NF-κB Pathway

doi: 10.1177/0963689719890597

Figure Lengend Snippet: Effects of cyclooxygenase 2 (COX2) on prostaglandin E2 (PGE2), PGE2 Receptor 2 (PTGER2) and p-nuclear factor-kappa B (NF-κB) p65 expression. (A) PGE2 release into the culture supernatants was measured by enzyme-linked immunosorbent assay. (B) The expression level of PTGER2 was estimated by quantitative polymerase chain reaction (qPCR). (C) The expression level of PTGER2, NF-κB p65, and p-NF-κB p65 and western blotting. The data are presented as the means±SDs.

Article Snippet: Membranes were blocked with 5% non-fat dry milk and were then incubated with primary antibodies specific for GAPDH (1:1000, Cell Signaling Technologies, cat. no. 2118), COX2 (1:1000, Cell Signaling Technologies, cat. no. 12282), p-NF-κB p65 (1:1000, Cell Signaling Technologies, cat. no. 3031), NF-κB p65 (1:1000, Cell Signaling Technologies, cat. no. 8242), CYP19 (1:1000, Santa Cruz, Dallas, TX, USA, cat. no. sc-374176), C-MYC (1:1000, Cell Signaling Technologies, cat. no. 5605), STAT3 (1:1000, Cell Signaling Technologies, cat. no. 9139), p-STAT3 (1:2000, Cell Signaling Technologies, cat. no. 9145), MMP2 (1:1000, Cell Signaling Technologies, cat. no. 4022), MMP9 (1:1000, Cell Signaling Technologies, cat. no. 3852), and PTGER2 (1:400, Bosterbio Technologies, Pleasanton, CA, USA, cat. no. BM5194) overnight at 4°C.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cell Transplantation

Article Title: Cyclooxygenase 2 Promotes Proliferation and Invasion in Ovarian Cancer Cells via the PGE2/NF-κB Pathway

doi: 10.1177/0963689719890597

Figure Lengend Snippet: Prostaglandin E2 Receptor 2 (PTGER2) and nuclear factor-kappa B (NF-κB) inhibitors inhibit the cyclooxygenase 2 (COX2)-promoted proliferation and invasion of ovarian cancer cells. (A) Cell viability was measured by a CCK-8 assay. a Compared Lenti-COX2 with Lenti-COX2+AH6809 and compared Lenti-COX2 with Lenti-COX2+BAY11-7802 separately, p < 0.05. (B) and (C) Representative immunohistochemical (IHC) staining of SKOV3 and OVCAR3 cells conducted with an antibody against Ki67. Magnification: ×400. (D) and (E) SKOV3 and OVCAR3 cells were seeded in 24-well Transwell chambers coated with Matrigel and cultured for 24 hours. Cell invasion was estimated. (F) The expression levels of CYP19, C-MYC, STAT3, p-STAT3, matrix metalloproteinase 2 (MMP2), and MMP9 were estimated by western blot analysis. The data are presented as the means±SDs.

Article Snippet: Membranes were blocked with 5% non-fat dry milk and were then incubated with primary antibodies specific for GAPDH (1:1000, Cell Signaling Technologies, cat. no. 2118), COX2 (1:1000, Cell Signaling Technologies, cat. no. 12282), p-NF-κB p65 (1:1000, Cell Signaling Technologies, cat. no. 3031), NF-κB p65 (1:1000, Cell Signaling Technologies, cat. no. 8242), CYP19 (1:1000, Santa Cruz, Dallas, TX, USA, cat. no. sc-374176), C-MYC (1:1000, Cell Signaling Technologies, cat. no. 5605), STAT3 (1:1000, Cell Signaling Technologies, cat. no. 9139), p-STAT3 (1:2000, Cell Signaling Technologies, cat. no. 9145), MMP2 (1:1000, Cell Signaling Technologies, cat. no. 4022), MMP9 (1:1000, Cell Signaling Technologies, cat. no. 3852), and PTGER2 (1:400, Bosterbio Technologies, Pleasanton, CA, USA, cat. no. BM5194) overnight at 4°C.

Techniques: CCK-8 Assay, Immunohistochemical staining, Immunohistochemistry, Cell Culture, Expressing, Western Blot

Journal: PLoS ONE

Article Title: EpCAM-Independent Enrichment of Circulating Tumor Cells in Metastatic Breast Cancer

doi: 10.1371/journal.pone.0144535

Figure Lengend Snippet: 2.5x10 4 EpCAM pos cells (pool of MCF7, SKBR3, HCC1500 and ZR-75-1; upper panel) and EpCAM low/ neg cells (MDA-MB-231; lower panel) were incubated for cell adhesion experiments on glass substrates (NEXTERION slides AL) manually coated with anti-EpCAM [Ber-EP4], anti-Trop2, anti-CD49f (0.1 mg/ml each), collagen I (Col), hyaluronic acid (HA) and laminin (Lam) (0.2 mg/ml each). Cell adhesion was visualized by Coomassie; 20x magnification.

Article Snippet: The following antibodies were used for immunofluorescence staining and immobilization on surface-modified glass slides (NEXTERION, Schott Technical Glass Solutions GmbH, Jena, Germany) and/or on immunomagnetic beads: anti-pan Cytokeratin-FITC (clone C-11) (GeneTex, Irvine, US), anti-EpCAM (clone Ber-EP4) (Dianova GmbH, Hamburg, Germany), anti-EpCAM (clone VU1D9) (Cell Signaling Technology, Cambridge, UK).

Techniques: Incubation

Journal: PLoS ONE

Article Title: EpCAM-Independent Enrichment of Circulating Tumor Cells in Metastatic Breast Cancer

doi: 10.1371/journal.pone.0144535

Figure Lengend Snippet: (A) CTC count determined by EpCAM-enrichment and subsequent CK/DAPI stain in 29 blood samples of 25 patients (DIII and DIV). (B) Number of potential CTCs (top) and double positive events (bottom) within the respective EpCAM-depleted supernatants of the same blood samples; total event numbers (CK pos /CD45 neg and CK pos /CD45 pos ) represent the sum of all events identified after 2–6 immunomagnetic enrichments for each blood/patient sample.

Article Snippet: The following antibodies were used for immunofluorescence staining and immobilization on surface-modified glass slides (NEXTERION, Schott Technical Glass Solutions GmbH, Jena, Germany) and/or on immunomagnetic beads: anti-pan Cytokeratin-FITC (clone C-11) (GeneTex, Irvine, US), anti-EpCAM (clone Ber-EP4) (Dianova GmbH, Hamburg, Germany), anti-EpCAM (clone VU1D9) (Cell Signaling Technology, Cambridge, UK).

Techniques: Staining

Journal: PLoS ONE

Article Title: EpCAM-Independent Enrichment of Circulating Tumor Cells in Metastatic Breast Cancer

doi: 10.1371/journal.pone.0144535

Figure Lengend Snippet: (A) One EpCAM pos CTC was selected and identified via CellSearch and re-identified with the CellCelector for a positive DAPI and CK-PE (displayed in the TRITC channel), and a negative CD45 (Cy5 channel) stain. Three EpCAM neg CTCs (I, II, III) from the CellSearch EpCAM-depleted fraction of the same patient were enriched with CD44-Adembeads and stained for DAPI/pan-CK-FITC/CD45-AF647. Single cells were isolated via the CellCelector, the whole genomic material was amplified and (B) genome wide aCGH profiles were obtained confirming that EpCAM-independent enrichment captures malignant cells. Chromosomal regions highlighted in grey show common somatic copy number alterations, light blue areas represent different chromosomal aberrations between both CTC populations.

Article Snippet: The following antibodies were used for immunofluorescence staining and immobilization on surface-modified glass slides (NEXTERION, Schott Technical Glass Solutions GmbH, Jena, Germany) and/or on immunomagnetic beads: anti-pan Cytokeratin-FITC (clone C-11) (GeneTex, Irvine, US), anti-EpCAM (clone Ber-EP4) (Dianova GmbH, Hamburg, Germany), anti-EpCAM (clone VU1D9) (Cell Signaling Technology, Cambridge, UK).

Techniques: Staining, Isolation, Amplification, Genome Wide